Table 1.
Cell and tissue slice cryopreservation by vitrification.
| Sample | CPA Solution | Cryocarrier | Sample Size | Viability Evaluation | Outcome | Ref. |
|---|---|---|---|---|---|---|
| Ovine embryo | 15% EG + 15% DMSO + 0.5 M sucrose + 30% Ficoll 70 | Cryotop-Spatula | <0.1 µL | Embryo morphology Cell membrane integrity with propidium iodide |
79.7% viability | [119] |
| Cow blastocyst | 16% EG + 16% DMSO + 0.5 M sucrose | Fork | 0.5 µl | Immunostaining | 74% hatching rate with warming in straw | [120] |
| Donkey embryo | 15% EG + 15% DMSO + 0.5 M sucrose + 18% Ficoll 70 | Cryotop | <1 µL | Transrectal ultrasonography to track follicular activity and confirm ovulation Cell membrane integrity with propidium iodide and Hoechst 33342 |
70% viability | [121] |
| Bovine oocytes | 2% EG + 2% PG | Cryotop | <1 µL | Oocyte viability, nuclear maturation status, embryo development, and blastocyst quality | 15% blastocyst yield | [122] |
| Mouse embryo | 20% EG + 0.4 M sucrose + 24% Ficoll 70 | Cryotube | 5 μL drop | Developmental ability | 98.7% viability | [123] |
| Human ovarian tissue | 30% EG + 0.5% trehalose + 6% FBS | Cold solid-surface, Straw, drop size into LN2 | 100 µL | Histologic analysis | Higher morphological follicles by the method of drop size into LN2 | [124] |
| Ovine cumulus-oocyte complexes | VS1, VS2 | Plastic insemination straw | 250 µL | Morphological evaluation, Trypan blue staining | 54.5% viability, 16.8% ultrastructural changes |
[125] |
| Bovine oocyte | 15% EG + 15% DMSO + 1.0 M sucrose | Hollow fiber | Inner diameter 200 μm, wall thickness 15 μm | Acetolacmoid staining | 23% blastocyst yield | [126] |
| Human embryo | 15% EG + 15% DMSO + 0.5 M sucrose | Plastic blade | Thickness: 0.05 mm | Assessment of pregnancy by transvaginal ultrasound imaging | 100% viability in Blastocyst | [127] |
| Bovine oocytes | 7.5% EG + 7.5% DMSO | Silk fibroin sheet multilayer | 0.1 mm thickness, 0.7 mm width, 10 mm depth | Morphological survival rates | 23% blastocyst yield | [128] |
| Rabbit chondrocyte sheets | 20% EG + 20% DMSO + 0.5 M sucrose + 10% COOH-PLL | Sealable polyethylene bag and nylon meshes | 110 × 85 mm; film thickness: 0.063–0.064 mm | Trypan blue staining Histological examination Immunohistochemical staining |
91% viability | [129] |
| Human liver tissue | 4.7 M 1,2-propanediol | Honeycomb-like tray | Diameter: 1 cm, Thickness: 200–250 µm | Xenobiotics metabolism | 7-EC metabolism, 7-HC conjugation (Results are very unstable) |
[81] |
| Human ovarian tissue | 7.5% EG + 7.5% DMSO + 20% FBS + 13.5% EG + 13.5% DMSO + 0.5 mol/l sucrose |
Needle directly into LN2 | 1 mm3 | Histologic Analysis by H&E Ultrastructural evaluation using TEM TUNEL assay for detection of apoptosis Assessment of tissue damage using an LDH assay |
Higher viability in stroma cells and lower apoptotic primordial follicles | [130] |
| Human ovarian tissue | 10% DMSO + 10% EG | Direct cover vitrification (DCV) |
1 × 1 × 1 mm | Follicle examination using electron microscopy and TUNEL | Higher normal follicles and lower apoptotic cells | [131] |
| Ovine testicular tissues | 18% EG + 18% DMSO + 0.5 M trehalose | E. Vit (modified plastic straw) | 1 mm3 | Cell plasma membrane integrity | 73.6% viability | [132] |
| Human ovarian tissue | 2.62 mol/L DMSO + 2.6 mol/L acetamide + 1.31 mol/L PROH + 0.0075 mol/L PEG |
20 μL droplet into LN2 | 4 mm × 4 mm × 1.5 mm | Immunohistochemistry histology | Higher follicles growth | [133] |
| Cat testicular tissues | 15% EG + 20% glycerol + 0.5 M sucrose | Needle | 1–2 mm3 | Seminiferous tubule Morphology Mitochondrial activity Cell composition |
92.9% viability | [134] |
| Human ovarian tissue | 40% EG + 1 M sucrose + 30% ficoll 70 | Cryovial | 2 mm3 | Histological examination Molecular assessment Hormonal assay Immunocytochemistry |
95.5% viability | [135] |
| Mouse testicular tissues | 15% EG + 15% DMSO + 0.5 M sucrose | Metal grid | Tissue fragments (0.5–1 mm2) | Trypan blue staining Hematoxylin and eosin (H&E) staining Immunohistochemistry staining |
97.7% viability | [136] |
| Dog ovarian tissue | 15% EG + 7.5% DMSO + 0.5 M sucrose + 2.5% PVP | Needle | Diameter: 2 mm | Neutral red staining Histology Xenotransplantation assays |
94.5% follicular viability | [137] |
| Rat testicular tissues | 15% EG + 15% DMSO + 0.5 M sucrose | Inoculation loop | Pieces of approximately 3 mm |
Trypan blue staining histological evaluation | 84.8% viability | [138] |
| Rabbit trachea | 18% EG + 22% DMSO + 0.5 M sucrose | Cryotube | 0.5 cm × 0.5 cm | Morphological and ultrastructural assessment HE examination TUNEL assays TEM and SEM |
97% viability | [139] |
| Rat kidney tissue | VM3 (8.44 M) in VS4 buffer (7.5 M) | Cryovial | Diameter: 5 mm | ATP content Histological integrity |
Cortex: histomorphology (86%), ATP content (113%); medulla: histomorphology (79%), ATP content (68%) | [140] |
| Rat liver tissue | VM3 (8.44 M) in VS4 buffer (7.5 M) | Cryovial | Diameter: 8 mm | ATP content Histological integrity |
histomorphology (71%), ATP content (58%) | [140] |
| Human osteochondral dowel | 9.5% EG + 18% DMSO + 5.8% PG + 14.1% glycerol + 0.1 mg/mL chondroitin sulfate (CS) | 5 mL vial | Full thickness in 10 mm diameter | Membrane integrity Metabolic activity Histology Immunohistochemistry |
75.4% viability | [141] |
| Pig osteochondral dowel | 16.6% EG + 21.5% DMSO + 22% PG + 0.1 mg/mL CS | Conical tube | two diameter sizes (10.0 mm and 6.9 mm) with 10 mm thickness | Chondrocyte assessment with membrane integrity stain and the chondrocyte metabolic activity by Alamar Blue. | 60–80% viability | [142] |
VM3 and VS4 are commercial CPA, consisting of buffer components such as NaCl, NaHCO3, KCl, …, and CPAs such as DMSO, Formamide, and Ethylene glycol.