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Botany JoVE (Open Access) Citable · DOI

Cryopreservation of Preimplantation Embryos of Cattle, Sheep, and Goats

DOI: 10.3791/2764-v
What you'll learn
  • ✓ Evaluate preimplantation embryos for cryopreservation viability
  • ✓ Execute controlled embryo dehydration and loading into straws
  • ✓ Perform slow freezing and liquid nitrogen storage protocols
  • ✓ Thaw and transfer cryopreserved embryos to recipient animals
Protocol

Biopharma Insights Preimplantation embryos may be cryopreserved after placement into a hypertonic cryoprotective solution to cause cellular dehydration. After equilibration, ice crystal formation is induced in the solution surrounding the embryo. Further dehydration occurs as the embryo is slowly cooled to subzero temperatures before plunging into liquid nitrogen for storage.

Difficulty
advanced
Total time
~4–6 hours per embryo batch (freezing cycle); thawing ~20–30 min per batch
Model organism
Cattle, Sheep, Goat (preimplantation embryos)
Biosafety
BSL-1

Steps

1
Evaluate embryo suitability for cryopreservation

Assess morphological quality and developmental stage of preimplantation embryos to determine candidacy for freezing. Select only viable embryos meeting established quality criteria.

▶ 02:28
2
Wash embryos to reduce pathogen carriage

Rinse embryos through multiple media changes to minimize disease transmission risk before cryopreservation and transfer.

▶ 03:58
3
Dehydrate embryos and load into plastic straws

Equilibrate embryos in hypertonic cryoprotectant solution to induce cellular dehydration, then load into 0.25 ml plastic straws for freezing.

▶ 05:20
4
Freeze embryos to liquid nitrogen temperature

Induce controlled ice crystal formation in surrounding solution and perform slow cooling to subzero temperatures before plunging straws into liquid nitrogen for long-term storage.

▶ 07:11
5
Thaw cryopreserved embryos for transfer

Warm straws from liquid nitrogen storage and remove embryos from cryoprotectant through stepwise rehydration before embryo transfer.

▶ 08:53
6
Transfer thawed embryos to recipient females

Place viable thawed embryos into synchronized recipient animals and evaluate pregnancy establishment and offspring outcomes.

▶ 10:26
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