ESHRE recommendations on Good Practice in the IVF laboratory

ESHRE Good Practice in the IVF Lab Working Group; Gemma Arroyo; Amy Barrie; Giovanni Coticchio; Thomas Ebner; Jackson Kirkman-Brown; Nathalie Le Clef; Kersti Lundin; Cristina Magli; Marina Quesada Martinez; Maria José de los Santos Molina; Kelly Tilleman; Ioannis Sfontouris

doi:10.1093/humrep/deag096

. 2026 Jul 6;41(8):1245–1269. doi: 10.1093/humrep/deag096

ESHRE recommendations on Good Practice in the IVF laboratory^{^†}

ESHRE Good Practice in the IVF Lab Working Group , Gemma Arroyo ^1,^✉, Amy Barrie ², Giovanni Coticchio ³, Thomas Ebner ⁴, Jackson Kirkman-Brown ⁵, Nathalie Le Clef ⁶, Kersti Lundin ⁷, Cristina Magli ⁸, Marina Quesada Martinez ⁹, Maria José de los Santos Molina ¹⁰, Kelly Tilleman ¹¹, Ioannis Sfontouris ^12,^13,^✉

PMCID: PMC13429877 PMID: 42405499

Abstract

STUDY QUESTION

What is the current good practice in the IVF laboratory, based on the best available evidence in the literature, if available, and the expertise of the working group?

SUMMARY ANSWER

The updated ESHRE Recommendations on Good Practice in the IVF laboratory provides recommendations on all activities in the IVF laboratory.

WHAT IS KNOWN ALREADY?

A previous version of the Good Practice in the IVF laboratory guideline was published in 2015. The adoption of new techniques, the increasing complexity of procedures, and the introduction of a revised European legislation on the quality and safety of tissues and cells for human application pertinent to assisted reproduction together necessitated an update of the document.

STUDY DESIGN, SIZE, DURATION

This document was developed according to a predefined methodology for ESHRE Good Practice recommendations. The working group reviewed the document of 2015, and based on this assessment, each group member updated one or more sections. Recommendations are supported by data from the literature, if available, and the expertise of the working group and were discussed until consensus was reached within the working group.

PARTICIPANTS/MATERIALS, SETTING, METHODS

The working group included 10 members representing the ESHRE Special Interest Groups for Embryology, Safety and Quality, and Andrology, with different areas of expertise and representing different European countries and settings. Based on the available evidence and the expertise of the working group, recommendations were formulated. Following stakeholder review of the initial draft, the final version was approved by the working group and ultimately by the ESHRE Executive Committee.

MAIN RESULTS AND THE ROLE OF CHANCE

The ESHRE IVF labs working group updated the recommendations on the general organization of an IVF laboratory (staffing and direction, quality management, and laboratory safety) and on the specific aspects of the procedures performed in IVF laboratories (identification of patients and traceability of their reproductive cells, consumables, handling of biological material, oocyte retrieval, sperm preparation, insemination of oocytes, scoring for fertilization, embryo culture and transfer, embryo biopsy for pre-implantation genetic testing, cryopreservation, and emergency procedures). The section on embryo culture and transfer was split up, the section on sperm preparation was expanded to cover all general andrological procedures, and a new section on biopsy procedure was introduced.

LIMITATIONS, REASONS FOR CAUTION

Not all recommendations are supported by evidence. Other recommendations, published in legal documents, relevant and recent documents, manuals, and consensus papers, were taken into account when formulating the recommendations.

WIDER IMPLICATIONS OF THE FINDINGS

The guideline group is confident that this document will be helpful to directors and managers involved in the management and organization of IVF laboratories and also to embryologists and laboratory technicians performing daily tasks.

STUDY FUNDING/COMPETING INTEREST(S)

The guideline was developed by ESHRE, which funded the guideline meetings, literature searches, and dissemination of the guideline. The guideline group members did not receive any financial incentives; all work was provided voluntarily. G.C. reports consulting fees from Gedeon Richter and Cooper Surgical and was part of the working group of the ‘Guide to the quality and safety of tissues and cells for human application’ of the European Directorate for the Quality of Medicines and Healthcare, on behalf of the Council of Europe. T.E. reports consulting fees from Nexpring Health and Esco Medical and speaker’s fees from Nexpring Health and Esco Medical. J.K.-B. reports research grants from Gates Foundation and NIHR, consulting fees from Bayer, speakers fees from Merck, IBSA, Ferring and Cooper Surgical, and travel support from Merck, IBSA, Ferring, and Cooper Surgical. I.S. reports speaker’s fees from Vitrolife and Cooper Surgical. I.S. also declares being a member of ARCS Scientific Committee. The other authors disclosed no conflicts of interest.

DISCLAIMER

This guideline represents the views of ESHRE, which were achieved after careful consideration of the scientific evidence available at the time of preparation. In the absence of scientific evidence on certain aspects, a consensus between the relevant ESHRE stakeholders has been obtained.

Adherence to these clinical practice guidelines does not guarantee a successful or specific outcome nor does it establish a standard of care. Clinical practice guidelines do not replace the need for application of clinical judgment to each individual presentation nor variations based on locality and facility type.

ESHRE makes no warranty, express or implied, regarding the clinical practice guidelines and specifically excludes any warranties of merchantability and fitness for a particular use or purpose (full disclaimer available at www.eshre.eu/guidelines).

Keywords: ESHRE, guideline, good practice, IVF, ICSI, embryo, embryology, laboratory, andrology

Introduction

The ESHRE Recommendations on Good Practice in the IVF Laboratory serve as a comprehensive guide to all procedures performed within the IVF laboratory, aiming to promote the highest standards of safety, quality, and effectiveness in medically assisted reproduction (MAR). Originally published in 2008 and subsequently updated in 2015, these guidelines have evolved in response to ongoing scientific advancements, technological innovations, and the changing regulatory landscape.

Since their inception, the recommendations have been developed through extensive literature review and expert consensus, establishing themselves as a valuable resource for MAR laboratory professionals. The adoption of new procedures and technologies has led to significant improvements in laboratory outcomes, while the introduction of updated European legislation has necessitated further revision of IVF laboratory procedures to ensure compliance and best practice.

This latest edition brings together the expertise of the ESHRE working group, integrating the most recent scientific evidence, stakeholder feedback, and regulatory developments. The document systematically lists and describes all relevant laboratory procedures, supporting harmonization of practices across different settings and fostering continuous improvement in patient care.

By providing clear, evidence-based recommendations, these guidelines are intended to assist laboratory directors, embryologists, technicians, and all members of the IVF team in delivering high-quality, patient-centred care. The present updated recommendations aim to promote excellence, safety, and accountability, ensuring that IVF laboratories remain at the forefront of innovation and best practice in MAR.

This Good Practice Recommendations paper was published before the sixth edition of the European Directorate for the Quality of Medicines and Healthcare (EDQM) guidance was finalized. However, the ESHRE working group took expected changes in the EDQM guideline into account based on the stakeholder version of the sixth edition. The sixth edition will be aligned with the new regulation and therefore compliance with this guide will equal compliance with the regulation.

Materials and methods

The current document was developed according to the manual for development of ESHRE Good Practice Recommendations (Vermeulen et al., 2019).

A working group was composed of members of the ESHRE Special Interest Groups (SIG) Embryology, Safety and Quality, and Andrology, ensuring representation of different types of laboratory expertise and geographical balance, supported by a methodological expert (N.L.C.). In the first meetings, the working group reviewed the topics that were covered in the 2015 version of the guideline, and missing topics were identified. A literature search of PUBMED/MEDLINE and Cochrane library was performed. Papers published up to 13 January 2026 were included. All titles and abstracts were screened to identify relevant papers, for which full-text papers were collected and summarized. At working group meetings, the evidence and draft recommendations were presented by the assigned working group member and discussed until consensus was reached within the group. Most of the recommendations from the 2015 version have been revised to reflect the more recent evidence cited in the main text preceding the recommendations. They have been summarized and supplemented with expert opinions. Recommendations with references added alongside them, however, are derived from those references.

Abbreviations used throughout this article are listed in Supplementary Table S1.

The final draft was published on the ESHRE website between 3 February and 3 March 2026 for stakeholder review. In total, 406 comments from 36 reviewers were received and incorporated where relevant. The review report is available on www.eshre.eu/guidelines. The experts who participated in the stakeholder review are listed in Supplementary Table S2.

Results

Staffing and direction

Personnel are one of the most important factors of an IVF laboratory. An IVF laboratory is both a diagnostic and therapeutic laboratory service unit and an integral part of the process of assisted reproduction. Appropriate human and logistical resources should provide conditions that allow all laboratory tasks to be performed in a timely and safe manner to ensure patient safety and quality care. The number of laboratory staff should reflect the number and complexity of tasks assigned to the embryologists at the specific clinic (see more in “continuing education” section), including duties such as administration, training, education, quality management, witnessing, and communication. Leave after weekend rotas, parental leave, illness, etc. also needs consideration in the number of staff. A minimum of two qualified clinical embryologists is always recommended in every laboratory, irrespective of the size and workload.

The hierarchical laboratory organization depends on staff size. Larger facilities can delegate responsibilities to different staff levels, e.g. supervisors, clinical embryologists, laboratory technicians, and administrative personnel (Alpha Scientists in Reproductive Medicine, 2015).

Laboratory director

The laboratory should be directed by a person with officially recognized qualifications and expertise in clinical embryology and biological/medical sciences. In accordance with the ESHRE certification programme, this would include a higher academic degree (MD, MSc, PhD) with a minimum of 6 years of documented experience in human embryology and preferably attainment of the ESHRE senior clinical embryologist certification (Kovačič et al., 2020; ESHRE Working Group on Embryologist Training Analysis et al., 2023) or similar. The laboratory director should also participate in national and/or international CPD (Continuing Professional Development) programmes.

Laboratory directors should be able to evaluate and interpret the significance of medical and laboratory findings and communicate them to laboratory staff members, clinical colleagues, patients, and the public. They should proactively seek clinical and scientific updates, promote staff performance and wellbeing, and promote science by participating in clinical studies and research, where possible.

Laboratory director responsibilities include ensuring:
• Implementation of a quality management system (QMS), including trouble-shooting.
• Development and updating of standard operating procedures (SOPs).
• Selection and implementation of the most adequate materials and procedures to reach the highest standards in clinical IVF.
• Maintenance of safe and appropriate laboratory facilities and equipment according to European and/or national regulations.
• Implementation of a laboratory risk management and prevention policy.
• Sufficient numbers of laboratory staff members with the appropriate skills.
• A comprehensive orientation and introduction programme for all new staff members.
• Management of laboratory staff training and continual scientific and biomedical education.
• Implementation and review of key performance indicators (KPIs) for all laboratory procedures for quality control and quality assurance purposes.
• Involvement in inspections from authorities and internal/external audits.
• Reporting of clinical data and adverse events (biovigilance) according to European and/or national regulations.
• Where relevant, participation in authorization and application of Standards of quality and safety for substances of Human origin (SoHO) preparation processes.
• Where relevant, participation in approval of research projects by competent authorities.

The QMS should include at least the following:
• The provision for unique identification of patients and their reproductive cells and tissues.Traceability records, whether maintained electronically or in handwritten form, should be clearly readable, permanent, securely stored, and readily accessible. Data should be electronically captured and validated; manual transcription should be double-checked.In absence of regulation stating otherwise, these records should be maintained securely for 30 years; critical materials and equipment data should be retained for at least 10 years.
• A documentation system for dealing with non-conformities, emergencies, incidents, adverse events or reactions, and complaints. Non-compliances should be analysed, and appropriate corrective and/or preventive actions (CAPA) should be identified. Non-conformities should be discussed with the teams regularly to highlight learning points and reviewed at least annually with the laboratory management.
• Procedures to address the withdrawal of release or (temporary) restriction of reproductive material in case of a recall or serious adverse events/reactions.
• Procedures for control of performance:
◦ Ensure performance indicators (PIs) are: • Defined on a specific patient reference group • Based on national data or European registry data (e.g. European IVF-monitoring programme for ESHRE) or other clinical group policies • Objective and relevant • Regularly monitored and reviewed using appropriate statistics for comparison • Communicated to all staff • Useful to take appropriate corrective actions when required ◦ Regularly assess operator performance by either direct observation of procedural skills or individual PIs follow up to ensure competence, compliance, and consistency. ◦ Implement retraining when necessary (i.e. extended periods of absence (e.g. parental leave, sick leave).	(ESHRE Special Interest Group of Embryology and Alpha Scientists in Reproductive Medicine, 2017; ESHRE Clinic PI Working Group et al., 2021)
• Participation in Internal Quality Control and External Quality Assurance programmes, either commercial or in collaboration with other laboratories, at least for diagnostic semen laboratories. These records should be maintained and reviewed, including documentation of results and any corrective action.
• A risk-based change-control system to describe steps to be taken to plan, record, analyse, and implement changes in the IVF laboratory.
• Audit systems, both internal and external, to verify compliance of all procedures with SOPs and requirements. Any findings, corrective actions, and their effectiveness should be documented.
• A documented validation of all critical processes and the qualification of all critical equipment used in these processes.
• A documented risk assessment strategy to identify, evaluate, and mitigate risks to ensure the protection of donor, recipients, and children born from MAR. ◦ Depending on the scenarios, use appropriate risk management tools; some examples are: • Hazard Analysis and Critical Control Points • Root Cause Analysis • Failure Mode and (Critical) Effects Analysis • EuroGTPII tool • EDQM Microbiological Risk of Contamination Assessment tool ◦ Perform structured periodic QMS reviews to: • Evaluate the overall system of all critical QMS elements • Ensure consistency and appropriateness • Identify trends • Identify and evaluate risks ◦ Conduct a comprehensive review, ideally every 3 years.	(European Committee on Organ Transplantation, 2022, Chapter 3)

More specifically:
• Materials used in laboratory construction, painting, flooring, and furniture should be appropriate for clean room standards, thus, minimizing release of volatile organic compounds (VOC) and embryo toxicity.	(Anagnostopoulou et al., 2022)
• Laboratory access should be restricted to authorized personnel. Entry of non-specialist staff should be documented.
• A double door system for clean access of personnel and materials to the laboratory is recommended.
• Staff should be provided with a dedicated space for changing into laboratory clothing and washing hands (with non-toxic disinfectants) before entering the laboratory.
• The area for cleaning and sterilization of materials should be separate from the laboratory.
• If possible, systems connected to main water and drainage should be outside of the laboratory due to potential infection risks associated with standing water.
• Separate office space for administrative work should be available outside the laboratory.	(Barrese et al., 2014)
• Potential distractors, such as private cell phones, should not be used in the laboratory.	(de los Santos and Ruiz, 2013)

• To optimize environmental conditions, laboratory air should be subjected to high-efficiency particulate air (HEPA) and VOC control. Irrespective whether qualitative or quantitative analysis of VOCs is done, a trend analysis should be considered.	(European Commission, 2022)
• Positive pressure is recommended to minimize air contamination. Air supply should ensure a sufficient number of fresh air changes.	(Cairo Consensus Group, 2020)
• Procedures involving gamete or embryo manipulation should be performed in a controlled environment. Background and processing air quality should comply with national and European guidelines and should be regularly monitored.
• The level of air cleanliness in the IVF laboratory should be risk-based and assessed at least once a year in ‘at rest’ and ‘in operation’ states.
• The microbial contamination level of the work surfaces in the laboratory should be regularly determined. Frequency of controls and number of sampling locations should be based on a documented risk assessment.

• The laboratory should contain all essential items required for all procedures being performed in a number appropriate to the workload. Critical items of equipment should be in duplicate.
• The number of incubators is critical and should be based on the number of cycles and embryo culture duration. Gametes and embryos should be conveniently distributed across incubators to minimize door openings causing fluctuations of physical parameters. In this context, TL incubators are associated with lower disturbance of culture conditions. Bench-top incubators show faster recovery times than big-box incubators.	(ESHRE Working Group on Time-lapse Technology et al., 2020)
• Equipment should be adequate for optimal laboratory work, easy to disinfect, and kept clean to avoid contamination.
• Critical equipment should be validated to meet quality requirements and to guarantee quality assurance. This process usually is composed of design, installation, operational, and performance qualification (PQ). Measured parameters should be verified by calibrated probes and instruments.
• Equipment should be marked for conformity with European standards (CE-marked) for the intended use where this option is available.
• Gas cylinders should be located outside of the laboratory. There should be an automatic change-over system and sufficient cylinders stocked for immediate replacement. High-purity/medical grade gas and inline HEPA and VOC filters are highly recommended.
• Regular maintenance should be scheduled and documented for all critical equipment, at least according to suppliers’ recommendations.
• Heating devices, such as tube warmers or heated stages, should be in place to maintain the optimal temperature of media as well as reproductive cells and tissues during handling.
• Accepted ranges of use for all measured parameters should be determined and monitored. If measurements are out of range, corrections should be made, documented, and their effectiveness verified.
• For every item of equipment, the instruction manual and simplified instructions, where needed, should be easily available.
• Malfunctioning equipment should be labelled as ‘out-of-use’ to avoid its use by mistake.
• Critical items of equipment, including incubators and cryostorage tanks, should be continuously monitored and equipped with external alarm systems.	(Anagnostopoulou et al., 2022)
• A backup power system should be in place for critical equipment.

• Procedures to ensure personnel safety and prevent cross-contamination should be established, taking national and European safety regulations into consideration. Therefore:	(ESHRE Guideline Group on Viral Infection/Disease et al., 2021)
◦ Vaccination of all personnel against hepatitis B or other viral diseases, for which a vaccine is available, is strongly recommended.
◦ Patients should be screened for infectious diseases according to national and European regulations.	(ESHRE Guideline Group on Viral Infection/Disease et al., 2021)
◦ Staff should be informed when a viral-positive patient is to be treated and be aware of the risks of handling infected biological material, as well as the correct procedures of disposal. However, crude samples (such as follicular fluid and semen) and patients without serology at the time of donation (e.g. oncological patients who come for sperm freezing) should always be treated as potentially infectious.
◦ SOPs should be in place to manage eventualities where infection might take place, e.g. needlestick injuries.
• To ensure adequate safety measures, the treatment of viral-positive patients should be only performed in IVF laboratories with dedicated areas and equipment. Alternatively, such patient treatments could be allocated to specific time slots, providing sufficient temporal and spatial separation and provided processing of their biological materials is followed by a thorough disinfection of the allocated areas and equipment. Procedures to handle gametes from viral-positive patients are covered in the ESHRE Viral Infection/Disease Guideline.	(ESHRE Guideline Group on Viral Infection/Disease et al., 2021)
• Whenever biological material is imported into the IVF laboratory from another clinic, full screening results should be obtained in advance. If any transported material is viral-positive, a dedicated dry shipper may be needed, depending on European and national regulations.

Each ART laboratory should:
• Encourage use of standardized witnessing protocols across all procedures and personnel to minimize variability
• When manual witnessing is used, it should involve two qualified staff members (double eyewitness) performing independent verification rather than passive confirmation by application of a standardized verbal confirmation protocol (e.g. read-back technique) to reduce confirmation bias
• Any witnessing discrepancy should trigger a predefined escalation protocol, including immediate procedure halt, documentation, and supervisory review
• Work scheduling should minimize fatigue during high-risk procedures (e.g. ICSI, embryo transfer), as cognitive overload increases error probability
• Encourage a blame-free culture for reporting witnessing mismatches or electronic witnessing systems (EWS) anomalies.
• Use reported cases and near misses as training material for continuous improvement.
• When using EWS take into consideration: ◦ The importance of staff training to maintain critical thinking and manual verification skills, even with EWS in place ◦ Data security standards, ensuring protection against unauthorized access, data loss, or manipulation ◦ Maintaining a validated manual witnessing backup protocol in case of technical failure ◦ The creation of indicators to minimize mismatch tolerance and clear user alerts to prevent overlooked errors ◦ Performance of test alerts during the implementation of a EWS to verify the correct functioning of processes and notifications ◦ Reiteration among team members about the complementary role of EWS as it does not replace human vigilance ◦ Emphasis of the importance of proper data entry and accurate sample labelling ◦ The implementation of EWS audits for design flaws, unclear alerts, test alerts, or normalization of deviation (e.g. ignored warnings)	(Intra et al., 2016; Gupta et al., 2020; Alteri et al., 2025) (Intra et al., 2016; Holmes et al., 2021) (Rienzi et al., 2015, 2017a; Vujisic et al., 2024)

For laboratories using EWS, governance should include scheduled audits (e.g. quarterly) of:
• Mismatch/near-miss typology	(Holmes et al., 2021; Ifenatuoha et al., 2023)
• False-positive alert rates
• Workflow interruption points
• Staff response consistency

• All medical devices used in the collection, processing, storage, and application of SoHO materials should comply with the stringent safety and quality standards established by the CE/Medical Device Regulation (MDR) ((EU) 2017/745).
• In-house manufactured medical devices may be used by IVF laboratories; however, under the MDR ((EU) 2017/745), health institutions may only manufacture and apply such devices internally if a comprehensive set of regulatory conditions is met.
• It is strongly recommended that consumables lacking CE/MDR are tested with an appropriate Mouse Embryo Assay/Human Sperm Survival Assay bioassay before using the materials in the clinical setting. It is also advisable to confirm that the pH of new batches of all culture media fall within the laboratory’s established acceptable range before use.	(Nijs et al., 2009; Chronopoulou and Harper, 2015; Delaroche et al., 2020; Togola et al., 2021; Delaroche et al., 2024)

• All reagents, culture media, and consumables must be used within the manufacturer’s stated expiry date to guarantee their efficacy and safety.
• Using products beyond their validated shelf life may compromise the viability of gametes and embryos.
• Regular inventory checks and a robust stock management system can aid in ensuring that expired products are promptly removed from use.	(Malhotra et al., 2021; Vujisic et al., 2024)

• Using appropriately sized containers reduces the risk of contamination and degradation by limiting exposure to air and temperature fluctuations between first and last use. This practice helps maintain the integrity of the products throughout their usage period.	(Chronopoulou and Harper, 2015; Delaroche et al., 2020; Vujisic et al., 2024)
• Consumables and media should be supplied in packaging that minimizes the need for repeated openings.

• Fertilization assessment should be performed under high magnification (at least 200×), using an inverted microscope equipped with Hoffman or equivalent optics (or a suitable TL microscopy device), in order to verify PN number and morphology.
• All inseminated or injected oocytes should be examined for the presence of PN and polar bodies (first and second) at 16–17 h post insemination (time suggested in case of static observation) in both conventional IVF and ICSI cases. For conventional IVF, cumulus cells should be removed, and fertilized oocytes transferred into new dishes containing pre-equilibrated culture medium or new clean drops of the same dish. The use of 2PN zygotes is the standard of practice. However, as reported from the Istanbul Consensus Update (Table 1), the use of specific categories of atypically fertilized zygotes may be considered.	(Working Group on the update of the ESHRE/ALPHA Istanbul Consensus et al., 2025)
• By static observation, PN may not be seen at fertilization checks, and yet normal embryo development can occur. This may be explained by TL technology data, which show that a significant proportion of 2PN zygotes undergo PN breakdown at earlier times than the above-recommended fertilization check interval. In such cases, the presence of the second polar body should accompany 2PN fertilization and therefore be used as a scoring criterion. While these zygotes may be incorrectly categorized as 0PN, if cultured, they may produce normal laboratory and clinical outcomes. Therefore, the term unfertilized or ‘0PN’ should not be used in these cases. Instead, ‘PN not observed’ may be a more suitable alternative for zygotes undergoing normal development without confirmation of fertilization.	(Working Group on the update of the ESHRE/ALPHA Istanbul Consensus et al., 2025)
• Preliminary preimplantation genetic testing for aneuploidy (PGT-A) data suggest that a significant proportion of 1PN zygotes may be biparental diploid. In addition, a growing number of studies have reported normal live births from 1PN zygotes derived from both ICSI and IVF cycles. Collectively, this evidence supports cautious clinical use of 1PN zygotes, combining blastocyst culture and, if available, PGT-A technology appropriate for biparental diploidy assessment. 2PN zygotes with one extra micropronucleus (2.1PN) are relatively rare. However, they also may have a diploid genotype and lead to apparently normal live births. Their clinical use may be considered, especially if associated with PGT-A technology. In general, the possible clinical use of 1PN and 2.1PN zygotes should be discussed with the clinical team and the patient and governed by an internally approved policy. The clinical use of zygotes with atypical pronuclear patterns should be fully traceable.	(Capalbo et al., 2024; Working Group on the update of the ESHRE/ALPHA Istanbul Consensus et al., 2025)

Pronuclear pattern	Laboratory recommendations	Clinical use without PGT-A technology appropriate for biparental diploidy assessment
PN not observed but presence of a second polar body (often incorrectly defined as ‘0PN’)	Culture to blastocyst stage to discriminate from rare non-fertilized oocytes undergoing early abortive cleavage (Coticchio et al., 2025).	Yes
1PN	Culture to blastocyst stage and, if available, PGT-A technology appropriate for biparental diploidy assessment	Yes, with caution, after discussion with the clinical team and the patients and governed by an internally approved policy.
2.1PN (2PN with a small micropronucleus)	Culture to blastocyst stage AND PGT-A technology appropriate for biparental diploidy assessment (Girardi et al., 2024).	Yes, with caution, after discussion with the clinical team and the patients and governed by an internally approved policy.
3PN	Culture to blastocyst stage AND PGT-A technology appropriate for biparental diploidy assessment	Not recommended as a routine practice, but admissible in pilot clinical studies

• Temperature of storage rooms should comply with the manufacturer’s specifications of the products.
• Storage conditions (e.g. refrigerated, frozen, room temperature) should be continuously monitored and documented.
• Refrigeration units that meet the specified temperature requirements should be available and monitored for the storage of media and reagents.
• Repeated exposure to temperature fluctuations during handling and storage should be avoided to preserve product integrity, as suboptimal storage conditions can lead to the degradation of media components, potentially impacting embryo development.	(Cairo Consensus Group, 2020; Vujisic et al., 2024)

Laboratories should:
• Record key data such as the batch number (lot number), date of entry into inventory, and expiration date to ensure traceability and prevent the use of expired materials.
• First-Expiry–First-Out inventory system should be implemented to ensure materials with the shortest remaining shelf life are prioritized for use.
• New batches of culture media or critical consumables should be quarantined upon receipt and released only after verification of documentation (e.g. certificate of analysis, lot validation).
• Where applicable, laboratories should perform internal validation or quality control testing when introducing new media or reagent lots to detect potential performance variability.
• A written policy should define acceptable in-use durations for opened media bottles, oil overlays, prepared dishes, and pre-equilibrated media.
• Expired or compromised materials should be clearly labelled as ‘out-of-use’ and removed promptly. Disposal should be documented according to laboratory waste management procedures.
• A minimum stock threshold should be defined for critical consumables to prevent last-minute use of near-expiry materials due to supply shortages.
• Where feasible, digital inventory systems should be used to automate expiry alerts and reduce human oversight errors.
• Implement standardized labelling and identification protocols to further enhance the safety and efficiency of laboratory operations	(Vujisic et al., 2024)
• Conduct regular risk assessments to ensure that all consumables and media are clearly labelled and easily identifiable; this minimizes the possibility of misuse and supports traceability and accountability in laboratory procedures.

• Handling of biological material should be easy, simple, and effective and should preferably be performed in laminar flow hoods equipped with heating stages and pre-warmed heating blocks, using aseptic techniques at all times.
• Measures should be taken to ensure that oocytes and embryos are always maintained at the appropriate temperature, pH, and osmolality during culture and handling. Exposure to light, toxic substances, or harmful radiation should be minimized.
• Traceability should be confirmed at all times (see Identification for patients and traceability of their reproductive cells section).
• MOPS/HEPES are preferably not used for oocyte or embryo culture. MOPS or HEPES (or similarly buffered solution) should be used if pH cannot be maintained during longer manipulations (e.g. denudation or ICSI). Appropriately buffered medium (bicarbonate, HEPES, or MOPS) should be selected based on the expected time taken for manipulation or culture as well as gas availability.	(Morgia et al., 2006; Mendola et al., 2024)
• Efforts should be made to reduce mechanical and shear stress on oocytes and embryos as far as possible through use of appropriately sized pipettes and minimizing handling.
• Oil overlay should be used for dishes whenever deemed appropriate ensuring appropriate volume and viscosity to reduce fluctuations in pH, temperature, and osmolality.
• Culture media and oil overlays should be adequately pre-equilibrated under appropriate gas conditions prior to use, according to manufacturer recommendations and internal validation protocols.
• Warming of pipettes and catheters for gamete or embryo handling is ineffective as the temperature loss is rapid when these consumables are removed from the source of heat.
• Reduce blue light exposure as much as possible.	(Cairo Consensus Group, 2020)
• Pipetting should be undertaken using standard hand pipettes. Mouth pipetting is not recommended.

• Temperature of heated stages and incubators should be regularly checked, since even slight changes (±1°C) can have a considerable effect upon any observed or measured sperm motility parameters.
• Semen samples should be collected into sterile, plastic containers (at least sperm-toxicity tested). The use of spermicidal condoms, creams, or lubricants should be avoided.	(Walker et al., 2024)
• All containers used throughout the process to hold sperm or semen should be clearly labelled with the patient’s identifiers. Records should also be kept of the type of container used and batch. The use of medication, fever during the previous months, abstinence, and completeness of the ejaculate collection should be documented.
• Collection should preferably be performed in a room near to the laboratory. If semen collection is performed outside the clinic premises, the patient needs to follow written instructions about handling the sample. After collection, the sample should be delivered to the laboratory as soon as possible, and within 60 min, avoiding extreme temperatures (<20°C and >37°C).
• An identity check should always be performed upon receiving a sample from a patient. Each laboratory should have in place a protocol to ensure the patient’s identification. The patient should verbally confirm their name and date of birth with the person receiving the sample from them and sign or electronically confirm that the sample is theirs.
• All semen or sperm handling steps should include appropriate laboratory witnessing and label checks; this is mandatory for the chain of custody. The time and place of collection and the time interval between collection and analysis/preparation should be noted for semen, as should times for each subsequent processing step.
• Sperm analysis and preparation should start as soon as possible when therapeutic use is intended (within 30 min if possible), as prolonged sperm exposure to seminal plasma may affect subsequent sperm quality. The seminal plasma osmolality also changes over time making sperm more prone to osmotic shock when being moved to media.	(Holmes et al., 2020)
• The following data on sperm preparation should be documented: ◦ sample origin (ejaculate/epididymal/testicular, donor/partner, fresh/frozen) ◦ preparation method and media used ◦ pre- and post-preparation sperm parameters
• An appropriate sperm preparation method should be chosen according to the characteristics and origin of individual samples. The swim-up technique and discontinuous density-gradient centrifugation are historically the most frequently used and widely accepted. Any preparation method including alternative methods should be validated by the clinic to determine the optimum sperm parameters required. This validation should include statistics relating to fertilization rates. Additional monitoring should be in place to ensure the selected method does not negatively impact the later pregnancy, miscarriage, or live birth rates (LBRs) (as these are also known to be independently influenced by sperm factors).	(West et al., 2022)
• In case of azoospermia on the day of oocyte retrieval and in the absence of a back-up sample, alternative sperm retrieval procedures or oocyte cryopreservation should be considered. Similarly, if an ejaculate cannot be produced on the day of treatment, potential interventions such as vibro-ejaculation may also be considered in addition to the aforementioned options.
• Where the ejaculate is below 1.4 ml and has a normal pH value (≥7.2), or totally absent when ‘dry orgasm’ occurs, then (partial) retrograde ejaculation may be suspected and evaluation completed in the diagnostic stages. If more sperm are required than available in the case of partial or total retrograde ejaculation, a method of sperm retrieval with bicarbonate can be used (see Supplementary Data File S1). NOTE: Potential treatments are available to reverse the retrograde ejaculation phenotype but are beyond the scope of this Good Practice Recommendations paper. NOTE: More complex methods of maintaining urine osmolality alongside pH have been suggested, but these have not as yet entered routine use or been widely tested. Older techniques involving catheterization of the bladder also lack robust evidence of additional benefit and require more invasive care so they are not recommended for routine use.	(World Health Organization, 2021) (Crich and Jequier, 1978) (Okada et al., 1998; European Association of Urology Guidelines, 2025) (Hotchkiss et al., 1954; Aust et al., 2008; Jefferys et al., 2012)
• Sperm obtained from surgical retrieval (PESA, TESA, TESE) require specific training to process.

• Temperature of heated stages and incubators, and the CO₂ and O₂ concentrations in incubators should be regularly checked.
• Before starting the procedure, the embryologist should verify that the patients have signed the corresponding informed consent and that the virological tests are valid.
• An identity check before oocyte retrieval is mandatory. Each laboratory should have in place a protocol to ensure the patient’s identification. The embryologist in charge of the procedure should check the patient’s name and date of birth. The identity check should include the labelling of dishes and tubes in contact with the collected specimens and should be documented. The identity checks should also be verified by a witness (double witnessing) or an EWS.
• Appropriate equipment should be in place to maintain oocytes close to 37°C. Flushing medium, collection tubes, and dishes for identifying oocytes should be pre-warmed. Media buffered to keep an appropriate pH in air should be used under strict temperature control, as temperature changes in buffered media can make pH conditions unstable, with negative consequences for oocytes. Alternatively, the procedure should be performed using a gassed chamber or in media overlaid with oil.	(Gatimel et al., 2020; Agarwal et al., 2022; Sciorio and Rinaudo, 2023)
• Follicular aspirates should be checked for the presence of oocytes using a stereomicroscope and heated stage, usually at 8–60× magnification. Exposure of oocytes to light should be minimized.	(Bódis et al., 2020)
• The time between oocyte retrieval and start of culture should be minimal.
• Timing of retrieval, number of collected oocytes, operator, and witness should be documented. A record should also be kept of the lot and type of disposables and media used.
• In some countries, oocyte retrieval is performed in one clinic and transported to another for fertilization and culture (so called transport IVF). Attention needs to be taken to keep the right temperature during transport, normally using a portable incubator.
• The oocyte retrieval rate is the ratio between the number of oocytes recovered and the number of ovarian follicles seen at ultrasound or aspirated. The expected range is 80–95% of follicles measured or aspirated in stimulated cycles. Values outside the expected range could be important to evaluate the ovarian stimulation protocol, the follicle assessment, and the oocyte retrieval technique.	(ESHRE Special Interest Group of Embryology and Alpha Scientists in Reproductive Medicine, 2017; ESHRE Working Group on Ultrasound in ART et al., 2019)

• The number of progressively motile sperm used for insemination should be sufficient to optimize the chance of normal fertilization. Traditionally, a progressively motile sperm concentration ranging between 0.1 and 0.5×10⁶/ml is used. Novel preparation methods, such as microfluidic separation, may result in reduced concentration but superior sperm quality. Novel methods should be validated by the clinic to determine the optimum sperm parameters required for fertilization.	(ESHRE Add-ons Working Group et al., 2023)
• An identity check before the oocyte insemination is mandatory. Each laboratory should have in place a protocol to ensure the identification of gametes at the time of the insemination procedure. This step should also be verified by a witness (double witnessing) or an EWS.
• Records should be kept of the time of insemination, the operator, and the concentration of progressively motile sperm used.
• Co-incubation of cumulus oocyte complexes and sperm is usually performed overnight, although a short co-incubation (2–4 h) can be considered following internal validation.	(Fan et al., 2023).

• Preparation of oocytes for ICSI.
◦ When removing cumulus cells from oocytes, hyaluronidase concentration and exposure should be kept to a minimum. ◦ In order to prevent oocyte damage, pipettes with appropriate lumen size should be used, and vigorous pipetting should be avoided. ◦ After enzymatic denudation, oocytes should be thoroughly washed to remove traces of hyaluronidase. ◦ The maturation stage of the oocytes should be recorded. ◦ Current evidence does not suggest that denudation should be performed at a specific time between oocyte recovery and ICSI.
• Preparation of oocytes for ICSI.
◦ An identity check before the oocyte insemination is mandatory. Each laboratory should have in place a protocol to ensure the identification of gametes at the time of the insemination procedure. This step should also be verified by a witness (double witnessing) or an EWS. ◦ Records should be kept of the injection time (start and end of the procedure) and the performing operator. ◦ The duration of sperm identification and immobilization followed by injection should be minimized. ◦ The number of oocytes transferred to the injection dish should relate to operator’s skills and sperm quality. ◦ During ICSI, the following points are important.
• Only mature oocytes should be injected
• Oocyte morphology should be recorded. Oocyte dysmorphisms such as large perivitelline space, localized granularity, and smooth endoplasmic reticulum aggregates may be associated with diminished clinical success. Giant oocytes should not be injected.	(Working Group on the update of the ESHRE/ALPHA Istanbul Consensus et al., 2025)
• Morphologically normal, motile sperm should be selected.
• Tail membrane breakage should be posterior to the midpiece and performed immediately before the injection of each individual oocyte.
• The polar body should be away from the injection site.
• Oolemma rupture should be assured prior to sperm injection.
• Volume of PVP/culture medium deposited in the oocyte with the sperm should be minimized.
◦ Appropriate temperature and pH should be maintained during injection. Viscous substances such as PVP can be used to facilitate sperm manipulation. In case of only immotile sperm cells, a non-invasive vitality test can be used to select viable sperm for injection. ◦ After injection, oocytes should be washed prior to culture.
• Artificial oocyte activation is currently not recommended for routine clinical use. It is, however, recommended for cases of previous complete activation failure (0% 2 pronuclei (PN)), very low fertilization (<30%), or globozoospermia.	(ESHRE Add-ons Working Group et al., 2023)

• A culture medium designed for embryo development should be used. There is currently insufficient evidence to recommend either sequential or single-step media as being superior for the culture of embryos to the blastocyst stage, both in terms of ongoing pregnancy rate, LBR, and neonatal outcome. In any case, media should be used according to the manufacturer’s recommendations.	(Sfontouris et al., 2016; Dieamant et al., 2017; Sacha et al., 2022; Sonigo et al., 2024)
• Oil overlay minimizes changes to temperature, pH, and osmolality. Its use reduces but does not eliminate evaporation and consequent changes in osmolality or fluctuations in temperature and pH. These changes depend on several factors, including microdrop size, type of culture dishes, incubator atmosphere, and intrinsic oil properties. Testing of oils with different viscosities indicates differences in protection against changes in osmolality or pH depending on the oil viscosity. There is, however, no solid evidence that this also results in differences in laboratory KPIs as well as clinical outcomes Oil for use in IVF culture systems must be tested to detect embryo toxicity. Storage and handling are critical and must follow the manufacturer’s recommendations	(Korakaki et al., 2020; Mestres et al., 2021; Scarica et al., 2022; Bossi et al., 2023) (Mestres et al., 2022; Murray et al., 2025; Watanabe et al., 2025) (Scarica et al., 2022)
• The type and number of incubators should be appropriate to the workload.
• For traceability purposes, single embryo culture is advisable. If group culture is used, the size of the droplet and the number of embryos per droplet should be considered and validated.
• To limit the damage caused by oxidative stress, low oxygen concentration (∼5%) should be used.	(Gardner, 2016; Herbemont et al., 2021, Slatinšek et al., 2026)
• Bacterial contamination of embryo culture is an uncommon event, with a reported incidence of <1%. When it does happen, the standard approach involves serial washing of the embryos in medium supplemented with high-dose antibiotics in an effort to clear microorganisms adhering to their surface. Removal of the zona pellucida (ZP) has also been suggested as a means of more effectively rescuing contaminated embryos. Following repeated washings, embryos may be cryopreserved while awaiting microbiological analysis of the contaminated medium, allowing the feasibility of subsequent transfer to be assessed on the basis of the contaminating organism and microbial load. Given the scarcity of published data on contamination in IVF, no formal recommendations can be formulated for such cases at present. Nonetheless, successful pregnancies have been reported following the strategies outlined above.	(Li et al., 2022; Vaduva et al., 2022; He et al., 2024; Sterckx et al., 2026)

• For the transfer procedure, the patient records should include: ◦ Date and time of embryo transfer ◦ Name of the operator ◦ Name of the practitioner performing the transfer ◦ Number, developmental stage, and quality of embryo(s) at the time of transfer ◦ Type of catheter used ◦ Fate of supernumerary embryos, when available ◦ Details about the procedure, e.g. presence of blood, retained embryo(s).
• It is recommended that the room in which the embryo transfer is performed be in close proximity to the laboratory. If the laboratory is some distance from the embryo transfer room, arrangements should be made to maintain temperature and pH whilst transporting embryos.	(Macklon et al., 2021; Gurner et al., 2024)
• A double identity check of the patient, the patient file, and the culture dish(es) is mandatory immediately before the transfer. This patient identity check should be documented.
• Catheter loading and technical aspects of the ET are described in D’Angelo et al. (2022). In summary:	(D’Angelo et al., 2022)
◦ There is insufficient evidence to suggest that flushing the loading catheter before transfer eliminates potential toxic agents benefiting clinical outcomes.	(Maldonado Rosas et al., 2022)
◦ There are few references in favour of performing an afterload (double step) versus direct (single step) catheter techniques.	(Sallam et al., 2022; Cirillo et al., 2023)
◦ There are few references in favour of the softness of the catheter or in relation to the material of the catheter used.	(Ebner et al., 2001; Practice Committee of the American Society for Reproductive Medicine, 2017)
◦ There is insufficient evidence to suggest the superiority of the air–fluid or fluid-only methods during embryo loading, and echogenic catheters available nowadays are easily visible by echography.	(Ebner et al., 2001; Abou-Setta et al., 2007; D’Angelo et al., 2022)
◦ Loading the catheter directly from the culture media under oil versus loading from a transfer dish without oil presents no differences in pregnancy rates.	(Halvaei et al., 2013)
◦ The use of specific transfer media enriched with adherence compound as hyaluronic acid has been demonstrated to be beneficial to the LBR in fresh transfers, although no effect has been demonstrated in FET. Due to the high variability of the studies and the increased multiple PR analysed, the Good Practice Recommendations on add-ons in reproductive medicine recommends their use and the need of further studies.	(Heymann et al., 2020; Yung et al., 2021; ESHRE Add-ons Working Group et al., 2023)
◦ The use of a small volume of medium (10–20 µl) is recommended. Other studies increased the transfer volume (to 35–45 µl) with controversial results in clinical outcomes.	(Ebner et al., 2001; Montag et al., 2002; Omidi et al., 2015; Sigalos et al., 2018)
• When ET is completed, both the outer and inner catheters have to be checked for possible retained embryo(s). The embryo should be re-transferred immediately, preferably with a new catheter, after embryo retention.	(ASRM, 2017; D’Angelo et al., 2022)

• Different cryopreservation approaches, including slow freezing and vitrification, can be used according to the type of biological material.
◦ For sperm, slow freezing (passive or controlled) is still the method of choice for ejaculated sperm. However, there are increasing improvements in vitrification options, particularly for low cell numbers, that, after careful evaluation, could be used as effective alternatives. ◦ For oocytes, pronuclear and cleavage-stage embryos, and blastocysts, vitrification is the more effective method and is recommended. ◦ For ovarian tissues, both slow freezing and vitrification are used. Recent studies indicate comparable results regarding follicular viability and the proportion of intact primordial follicles. However, cryopreservation protocols differ widely, and comparative data on results after re-transplantation are lacking. Vitrification of ovarian tissue may therefore still be considered innovative. ◦ For cryopreservation of testicular tissue from prepubertal boys, not expected to contain sperm, slow freezing is recommended (passive or controlled).Where tissue potentially may contain sperm, it is recommended to analyse the testicular sample to determine if sperm is present. If sperm are identified, a protocol for sperm cryopreservation should be favoured over testicular tissue cryopreservation. If there are no sperm present, testicular tissue cryopreservation should be favoured. Alternatively, part of the tissue could be cryopreserved using a protocol aimed at preserving spermatogonia and the other part to preserve sperm. ◦ Semi-automated or automated vitrification has been shown to have similar survival, embryo development, and pregnancy results as manual vitrification but still needs to be further evaluated in (large) randomized studies. ◦ Supernumerary embryos should be individually cryopreserved, or may be donated to research or training, or discarded, according to their quality, the patient’s wishes, and national legislation.	(Ozimic et al., 2023) (Rienzi et al., 2017b; Golakov et al., 2018; Hajek et al., 2021) (ESHRE Guideline Group on Female Fertility Preservation et al., 2020; Sugishita et al., 2021; Antonouli et al., 2023; Cariati et al., 2023; Hončová, 2023; Pantos et al., 2024; Kong et al., 2025) (ESHRE FP for Boys Working Group et al., 2025) (Miwa et al., 2020; Gatimel et al., 2021; Hajek et al., 2021)
• Any risk of transmission of infection via LN2 should be minimized:
◦ The risk of contamination from LN2 or from other biological samples has been considered to be negligible; however, concern is being raised regarding possible risk for viral contamination (see also Laboratory safety section). Closed devices have shown to have similar cryosurvival, embryo utilization, and pregnancy rates as open devices. Laboratories should make decisions based upon risk analysis and regulations in place. ◦ Specimens from sero-positive patients should be stored in high-security closed devices. Dedicated tanks are recommended.	(De Munck et al., 2016; Cai et al., 2018; Pomeroy and Schiewe, 2020; De Santis et al., 2021; Porcu et al., 2021; Sugishita et al., 2021; Vajta et al., 2022) (ESHRE Guideline Group on Viral Infection/Disease et al., 2021)
• At cryopreservation, documentation on biological material should include: ◦ Patient consent form ◦ Labelling of devices ◦ Cryopreservation method ◦ Date and time of cryopreservation ◦ Operator ◦ Sperm quality ◦ Embryo quality and stage of development ◦ Number of oocytes or embryos per device ◦ Number of devices stored per patient ◦ Location of stored samples (tank, canister) ◦ Unique code or SEC codes when applicable.	(European Committee on Organ Transplantation, 2026)
• Cryo-devices should be clearly and permanently labelled with reference to patient details, treatment number, and/or a unique identification code.
• A periodic inventory of the contents of the cryobank is recommended, including cross-referencing contents with storage records. In some EU countries, national laws regulate a maximum legal storage period for gametes and/or embryos.
• At thawing (warming), documentation on biological material should include: ◦ Thawing method ◦ Date and time of thawing ◦ Operator ◦ Post-thawing sample survival and quality.
• A double-check of patient identity and embryo number is recommended in the following steps: transfer of samples into labelled cryo-dishes, loading of the labelled device, deposition in the cryobank, and removal from the cryobank. This patient identity check should be documented.
• During storage, handling, and transport of cryopreserved material, care should be taken to maintain adequate and safe conditions. Temperatures should never rise above −140°C. During storage and transport, temperature loggers or indicators should be used.	(European Committee on Organ Transplantation, 2026)
• In case of transport of cryopreserved SoHO materials, it needs to be clearly documented which centre is responsible from release to receipt. The distribution to another establishment should be authorized by a responsible person and restricted to authorized entities. Written agreements should be in place.Reports should be established, and include: ◦ Name and identification of distributing and receiving parties ◦ Name and identification of transporting party ◦ Type of sample and packaging ◦ Time and date of distribution and delivery ◦ Patient identification codes ◦ Any possible incidents occurring during the transport	(European Committee on Organ Transplantation, 2026)

PERMALINK

ESHRE recommendations on Good Practice in the IVF laboratory†

Gemma Arroyo

Amy Barrie

Giovanni Coticchio

Thomas Ebner

Jackson Kirkman-Brown

Nathalie Le Clef

Kersti Lundin

Cristina Magli

Marina Quesada Martinez

Maria José de los Santos Molina

Kelly Tilleman

Ioannis Sfontouris

Abstract

STUDY QUESTION

SUMMARY ANSWER

WHAT IS KNOWN ALREADY?

STUDY DESIGN, SIZE, DURATION

PARTICIPANTS/MATERIALS, SETTING, METHODS

MAIN RESULTS AND THE ROLE OF CHANCE

LIMITATIONS, REASONS FOR CAUTION

WIDER IMPLICATIONS OF THE FINDINGS

STUDY FUNDING/COMPETING INTEREST(S)

DISCLAIMER

Introduction

Materials and methods

Results

Staffing and direction

Laboratory director

Laboratory supervisors/manager

Clinical embryologists

Continuing education

Number of staff

Quality management

Laboratory safety

Laboratory design

Laboratory air quality

Laboratory equipment

Cryopreservation facilities and material

Infectious agents

Protective measures

Identification for patients and traceability of their reproductive cells

Unique identification code

Witnessing protocols

Consumables

Critical reagents and materials

Consumable delivery entry control

Use of sterile, single-use disposables

Expiry dates and shelf life management2

Appropriate packaging size

Storage conditions

Stock management system

Handling of biological material

General andrology procedures

Oocyte retrieval and processing

Insemination of oocytes

Conventional IVF

ICSI procedure

Scoring for fertilization

Embryo culture

Different approaches or culture systems can be used in order to optimize embryo development

Embryo quality assessment

Biopsy procedure

Method of fertilization

Embryo culture

Embryonic stage of biopsy

Method of zona opening

Technique of cell removal

Tubing

Rebiopsy and refreeze

Embryo transfer

Cryopreservation

Contingency and emergency plan

Contingency plan

Emergency plan

Discussion

Table 1.

Supplementary Material

Acknowledgements

ESHRE recommendations on Good Practice in the IVF laboratory^{^†}

A contingency plan for the IVF laboratory contains at least the following items (please find a comprehensive list in the Cairo Consensus: • Alarm system ◦ The laboratory should have an alarm system to notify staff of malfunctioning or failure of critical equipment or occurrence of adverse environmental conditions in the laboratory.	(Cairo Consensus Group and Alpha Scientists in Reproductive Medicine, 2025)
• Facilities ◦ Electricity: loss of electrical power should be compensated by generators or uninterrupted power supply (UPS) systems. It could be useful to have a separate UPS unit specifically for the lab in case the hospital system fails or needs this power to keep operating theatres running and live support for patients. ◦ LN2: in case of failure of automatic supply lines, tanks should be filled manually. A reserve LN2 tank should be available, and knowledge of how low the tanks can keep their temperature without being filled. ◦ Gases: have a strategy to minimize disruption in gas supply and have a backup plan in case it happens.
• Equipment ◦ In case of power failure, critical equipment should be prioritized. ◦ A second item of critical equipment should be available if the first item fails. All reserve equipment should be fully validated and ready for use. ◦ Back-up cooled freezers (−20°C) and refrigerators should be available.
• Contingency culture/freezing protocols ◦ Based on the remaining facilities in the laboratory, it could be necessary to prioritize vitrification of material to safeguard all the reproductive material.	(Sharma et al., 2024)
• Medical records ◦ Records to identify the ownership of human tissue should be kept on a secure web server.
• Staff ◦ Have sufficient and well-trained staff and have backups for key personnel
• Consumables ◦ Assess the amount of consumables needed to be able to execute contingency and emergency laboratory protocols. Evaluate the possibility to reach out to other suppliers if necessary.