|
Animal | Cryopreservation methods | Reference number |
|
Human | | |
| Semen was diluted using human sperm preservation medium, TEST-Yolk buffer, or glycerol, subjected to slow manual cooling in liquid nitrogen (LN2) vapour, and stored in LN2. | [47] |
| The procedure is based on the program of cooling speed doubling: from 20 to 5°C at 0.5°C/min; from 5 to 4°C at 1°C/min; from 4 to 3°C at 2°C/min; from 3 to 2°C at 4°C/min; from 2 to 1°C at 8°C/min; from 1 to −80°C at 10°C/min. After being held 10 minutes at the final temperature, −80°C then were transferred to LN2. | [62] |
| Normozoospermic, oligozoospermic, asthenozoospermic, and oligoasthenozoospermic semen samples were frozen in pellets on the surface of dry ice using a glycerol-based cryoprotectant with egg yolk. | [65] |
| Sperm cells were frozen or cold-shocked by lowering the temp. rapidly from 37 to 0°C on melting ice. | [66] |
| Sperm cells were frozen using the programme: from 20 to −4°C at 5°C/min; from −4 to −30°C at 10°C/min; from −30 to −140°C at 20°C/min and then were transferred for storage to LN2. | [67] |
Ram and goat | | |
Merino rams | Ejaculates were obtained by electrical stimulation. Semen was also collected from the caput and cauda epididymis. Spermatozoa were cold-shocked by placing tubes containing the semen held at 37°C into a bath at 0°C for 10 min. |
[20] |
Zandi rams | The diluted semen was cooled at 4 to 5°C for 2 h. Then the samples were placed into the LN2 vapor at a height of 4 cm above the liquid for 8 minutes, and then the straws were plunged into LN2. | [80] |
Goat | Epididymal sperms were cryopreserved using programmable freezer: from 30 ± 2° to 5°C at 0.25°C min, from 5 to −20°C at 5°C min, and from −20 to −100°C at 20°C min then transferred to LN2. | [81] |
Goat | Semen was frozen in pellet form on dry ice, and then plunged into LN2. | [82] |
Mahabadi bucks | Ejaculated semen samples were diluted and equilibrated at 5°C for 150 min. Samples were frozen in LN2 vapor, 4 cm above the L, for 7 min; subsequently the straws were plunged into the LN2 for storage. | [83] |
Blanca-Celtiberica buck | Diluted semen was cooled to 5°C for 2 h, diluted further, and held at 5°C for 2 h; another sample was cooled to 5°C for 4 h; both samples were frozen over N2 vapour for 10 min, 4 cm above N2 level, plunged, and stored in LN2. | [84] |
San Clemente bucks and Tennessee Myotonic buck | Semen aliquots were cooled on ice for 1 h before transfer to a 5°C refrigerator and kept for total of 1-2 h; then extender was added. Fully extended semen was equilibrated at 5°C for 2 h. Semen was frozen by suspending straws in N2 vapour for 10 min, then plunging into LN2. | [85] |
Saanen bucks | Semen samples were cryopreserved using programmable freezer using a fast freezing curve (from 25°C to 5°C at 0.25°C/min and from 5°C to −120°C at 20°C/min) that started at 28°C. After reaching a temperature of 5°C (~80 min), the straws were subjected to an equilibration time for 120 min. The freezing curve was implemented immediately after the equilibration time and was sustained until the temperature reached −120°C, then were placed in LN2. | [87] |
Spanish ibex (Capra pyrenaica) | The diluted sperm suspension was cooled to 13°C in a water bath, further cooled to 5°C over 1 h, kept for 2 more h, and frozen by placing in N2 vapour 5 cm above the surface of LN2 for 10 min before plunging into the LN2 . | [102] |
Bull | | |
Friesian bull | Semen was cooled to 5°C over 30 min, equilibrated for 6 h at 5°C, and frozen above the surface of LN2 (a temperature of −120°C was attained in 7 min). | [73] |
Holstein bulls | Semen was cooled to 5°C in 1 h; after 4 h it was pellet-frozen on solid CO2. | [74] |
Holstein, Jersey, and Guernsey | The semen was cooled immediately after collection to 15 to 20°C and held at this temperature for one-half to one hour; then diluted semen was cooled slowly to a storage temperature between 4 and 7°C. | [76] |
Prim Holstein | Electroejaculated semen samples were cooled from 34 to 4°C in 1.5 h, held for 2 h, and then descended in LN2. Rate: 4 to −10°C at 0. 7°C/10 s; −10 to −150°C at 7°C/10 s. | [77] |
Holstein | Fresh sperm samples were cooled to 4°C and held for 1 h (addition of diluent). Cryostraws were placed ~5 cm above the LN2 surface for 10 min and then put directly into the LN2 for storage | [78] |
Swiss brown bull | Diluted semen was cooled to 4-5°C over 2 h and then frozen by being placed into the LN2 vapor at a height of 4 cm above the liquid for 8 min; after that, the straws were plunged into LN2. | [79] |
Boar | | |
| The semen was cooled to 22°C over 2 h and further cooled to 5°C over 3 h by placing in a cold room. Samples were then frozen in LN2 vapors at 30°C/min from 5°C a final temperature of −70°C. Then straws were plunged directly into LN2. | [38] |
Norwegian Landrace and Duroc | The diluted semen was cooled at 15°C over 3 h and then cooled to 4°C over 2 h period. Semen was frozen in a controlled rate freezer for 9 min. The freezing chamber was precooled to −100°C. Immediately after the straws were transferred to the chamber it was warmed at 10°C/min to −70°C and held for 1 min before lowering the temperature to −120°C at 50°C /min. The straws were held at −120°C for 4 min before transferring to LN2. | [11] |
Landrace, Large White, and commercial hybrids | Semen was cooled at 15°C for 3 h and was further cooled in a programmable freezer to 5°C over 90 min; samples were cryopreserved using programmable freezer. The freezing chamber was precooled to −110°C. Immediately after loading the straws it was warmed at 45°C/min to −60°C and held for 1 min before lowering the temperature to −130°C at 20°C/min. The straws were plunged into LN2. | [41, 90] |
Marsupial | | |
Eastern grey kangaroo (Macropus giganteus) | Epididymal sperms were subjected to cold shock by rapid cooling. |
[91] |
Koala (Phascolarctos cinereus) |
Common wombat (Vombatus ursinus) |
Canine samples | | |
Blue fox (Alopex lagopus) | Semen was cryopreserved in different extenders; FA and sterols of plasma membrane were analysed. | [10] |
Red/silver fox (Vulpes vulpes) | Cooling at a moderate rate (2–5°C/min from 4-5°C to below the freezing point, i.e., from −7 to −15 or −20°C) and freezing at a rapid rate from −20 to −50 or −70°C. | [92, 94, 95] |
Dog | Cooling at a moderate rate (2–5°C/min from 4-5°C to below the freezing point, i.e., from −7 to −15 or −20°C) and freezing at a rapid rate from −20 to −50 or −70°C. | [94] |
Beagle and Golden Retriever | The samples were maintained at a temperature of 4°C for total 1 h 30 min. Then the canine semen was frozen in LN2 vapours (4 cm above the level of the LN2) at −110°C for 10 min then immersed vertically in LN2 for storage at −196°C. | [99] |
Elephant | | |
African Elephant (Loxodonta Africana) | Ejaculated semen samples were stored at −70°C or cryopreserved in LN2. |
[9] |
Asian Elephant (Elephas maximus) |
Stallion | | |
Andalusian stallions | The spermatozoa were slowly cooled to 4°C within 1 h and frozen horizontally in racks placed 4 cm above the surface of LN2 for 10 min, after which they were directly plunged in LN2. | [100, 101] |
Bats/flying fox | | |
Indian flying-fox (Pteropus giganteus) | Electroejaculated semen samples were cooled at 50°C/min to 4°C. |
[12] |
Variable flying-fox (Pteropus hypomelanus) |
Grey-headed flying-fox (Pteropus poliocephalus) |
Rodrigues flying-fox (Pteropus rodricensis) |
Large flying-fox (Pteropus vampyrus) | |
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