. 2016 Aug 5;33(10):1261–1272. doi: 10.1007/s10815-016-0774-5

Table 1.

Characteristics of the included studies

Study	Country	Enrollment	Eligibility criteria	Culture condition (single)	Culture condition (sequential)	Authors’ conclusions
Studies included in the quantitative analysis
Randomization of women
Macklon 2002	The Netherlands	Sep 1999 to Sep 2000	Indication for IVF	Rotterdam culture media (Group 1: without change on D3—Rotterdam monoculture system; Group 2: media changed on D3—Rotterdam serial culture system); culture in a closed incubator at 37 °C in 5 % CO₂	Vitrolife G1 until day 3 and Vitrolife G2 until day 5; culture in a closed incubator at 37 °C in 5 % CO₂	No significant differences in blastulation and implantation rates
Sfontouris 2015	Greece	Jan 2014 to Jun 2014	Age ≤ 40 years; at least 10 retrieved oocytes; fresh cycles with own eggs	Global medium, media was not changed on D3	Origio (ISM1/BlastAssist), media was changed on D3	Implantation and pregnancy rates, as well as blastocyst formation and embryo utilization rates were similar
Randomization of oocytes/zygotes
Alteri 2014	Italy	Sep 2014 to Dec 2014	Women aged ≤39 years	Unclear media, SAGE, media was not changed on D3; culture was in multi-gas incubators (Sanyo) at 37 °C, 5 % 0₂, 5.5 % CO₂	Unclear media, SAGE, media was changed on D3; culture was in multi-gas incubators (Sanyo) at 37 °C, 5 % 0₂, 5.5 % CO₂	Single media yielded a significant higher blastocyst formation and top-quality blastocyst rate
Donmez 2008	Turkey	NR	NR	Single-Step Medium (SSM), media was not changed on D3	Sequential complete P-1 Medium, media was changed on D3	Single media seems to be more effective on blastomere compaction and blastocyst development
Khoury 2012	USA	NR	NR	Global, media was changed on D3; Global medium was supplemented with Life Global Protein Supplement, overlaid with Global mineral Lite Oil	Quinn’s Advantage (QA) cleavage medium until day 3 and (QA) blastocyst medium until day 6; SAGE media was supplemented with SAGE SPS protein, overlaid with SAGE mineral oil	Single media increased blastocyst formation and number of embryos transferred and resulted in slightly improved clinical pregnancy rate and implantation rate
Koyner 2013	Panama	NR	First IVF treatment	SMS (Global), half renewal and half no renewal media on D3; culture was at 37 °C and 6 % CO₂	Sequential Media System—SQMS (SAGE) media was changed on D3; culture was at 37 °C and 6 % CO₂	The renew of 50 % of media during the embryo culture in SMS increase the embryo cell division and have better potential in achieving good pregnancy and implantation rate
Patrick 2013	North Carolina	NR	NR	Single media: not reported	Sequential media: not reported	Blastocyst formation was similar
Sepulveda 2009	Peru	Mar 2006 to Oct 2006	Oocytes donors age 19–34 years (not randomized); recipients (randomized)	Global medium, media was change D3; culture was at 5.6 % CO₂	Irvine Early Cleavage Medium (ECM) until day 3 and Multiblast (Irvine Scientific) until day 5–6; culture was at 5.6 % CO₂	A single medium was as good as or better than a sequential media system for human embryo culture from the zygote to blastocyst stage
Sfontouris 2014	Greece	Oct 2013 to Jan 2014	Age ≤ 40 years; ≥10 retrieved oocytes	Global medium, media was change D3	Origio (ISM1/BlastAssist), media was changed on D3	Culture in a single-step medium is associated with higher blastocyst formation rates, better blastocyst quality, and higher blastocyst utilization rates compared to sequential media
Summers 2013	London	NR	Age ≤ 38 years, first or second IVF cycle, ≥6 2PN	Global medium, media was change D3; culture was in Planar BT37 incubators (Planar) humidified at 37 ± 0.5 °C in an atmosphere of 6 % CO2/6 % O2/88 % N2	Quinn’s Advantage Cleavage medium (SAGE) until day 3 and Quinn’s Advantage Blastocyst medium until day 6; culture was in Planar BT37 incubators (Planar) humidified at 37 ± 0.5 °C in an atmosphere of 6 % CO2/6 % O2/88 % N2	No differences were noted in the rates of blastocyst development, inner cell mass (ICM), and trophectoderm (TE) scores in the two culture media
Werner 2016	New Jersey	Aug 2013 to Mar 2015	Age ≤ 42 years, FSH < 12 IU/L, AMH ≥ 1.2 ng/mL, and AFC ≥ 12, not requiring surgical sperm extraction	Continuous Single Culture (Irvine Scientific), media was not change D3; culture was in 5 % O₂	Quinn’s Advantage Cleavage Medium (SAGE) followed by Blast Assist (Origio), media was changed on D3; culture was in 5 % O₂	Blastocyst rate is greatest after culture in the sequential media. However, no difference exists in timing of blastulation, aneuploidy, or sustained implantation rates
Yang 2013	China	NR	Indication for preimplantation genetic screening	Single culture media—CSC, Irvine Scientific, not clear if renewal media on D3	Vitrolife G1 until day 3 and Vitrolife G2 until days 5 and 6	The key morphokinetical parameters and percentage of euploid blastocysts are similar
Studies not included in the quantitative analysis
Randomization of women
Koscinski 2014	France	NR	First and second IVF attempts with own eggs, age < 37 years, no indication for PGD	Global medium, media was not changed on D3	Sequential media Cook, media was changed on D3	More physiologic embryo development kinetics and higher rates of blastocyst were after culture in the sequential media
Scarica 2015	Italy	Sep 2013 to Sep 2014	NR	Single-step medium, media was not changed on D3; culture was in a bench\top!incubator (MINC) or in the Embryoscope (Unisense)	Embryo cultured, SAGE, media was changed on D3; culture was in a bench\top!incubator (MINC) or in the Embryoscope (Unisense)	Single medium culture system has an increased blastocyst developmental rate. No significant differences were found in aneuploidy rate and clinical outcomes
Randomization of oocytes/zygotes
Ciray 2012	Turkey	Mar 2011 to Dec 2011	Women who retrieved 2–6 MII, Age < 40 years, no indication for PGD	Single-Step Media, SSM, media was changed on D3; culture was in time-lapse incubator (Embryoscope ^TM) at 37 °C, 50₂, 6 % CO₂	Early Cleavage Media, ECM, until day 3 and MultiBlast Media, MB, until Day 6; culture was in time-lapse incubator (Embryoscope ^TM) at 37 °C, 5 % 0₂, 6 % CO₂	The overall clinical and embryological parameters remain similar
Hardarson 2015	Four centers, two in Sweden and two in the USA	Sep 2013 to Feb 2014	Age ≤ 40 years, ≥ 6 2PN, not requiring surgical sperm extraction	Vitrolife (G-TL), media was not changed on D3; Culture was in time-lapse incubator at 37° C (6–7 % CO₂, 5 % O₂, and 89 % N₂)	Vitrolife G1 until day 3 and Vitrolife G2 until day 5–6; culture was in time-lapse incubator at 37 °C (6–7 % CO₂, 5 % O₂, and 89 % N₂)	Two media systems are equivalent in relation to blastocyst quality and rate
Reed 2009	USA	Jun 2008 to Dec 2008	≥2 2PN, no indication for PGD or PGS	Global medium (IVFonline), media was not changed on D3; Culture was at 37 °C and 6 % CO₂	Vitrolife G1 until day 3 and Vitrolife G2 until days 5 and 6; culture was at 37 °C and 6 % CO₂	There is no apparent advantage in using sequential media over a single medium
Schneider 2009	Brazil	NR	NR	Global medium (LifeGlobal), media was change D3	Vitrolife G1 until day 3 and Vitrolife G2 until day 5	Single medium is as effective as sequential medium for the development of human embryo to the blastocyst stage
Vansteenbrugge 2007	Belgium	NR	≥7 retrieved oocytes	Global medium, IVFonline, media was changed on D3; culture was in 6 % CO2 and 5 % O₂ at 37 °C	Vitrolife G1 until day 3 and Vitrolife G2 until day 5; culture was in 6 % CO₂ and 5 % O₂ at 37 °C	The ability of embryos cultured in the single medium to further development to blastocyst and implant is similar to embryos cultured in the sequential media
Velez de la Calle 2013	France	Oct 2012 to Mar 2013	NR	Continuous single culture—CSC from Irvine, media was not changed on D3; culture was in 6 % CO₂ at 37 °C	Ferticult from Fertipro until day 3 and CCM from Vitrolife until day 5; culture was in 6 % CO₂ at 37 °C	Single media resulted in more blastocysts, although clinical pregnancy rate was not significantly different