High-throughput and non-invasive method to vitrify porcine embryos
AssigneeThe Curators of the University of Missouri
InventorsRandall S. Prather, Rongfeng Li, Clifton N. Murphy, Lee Spate
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Abstract
The present invention provides a practical, non-invasive, and efficient method for cryopreservation of an in-vitro-produced porcine embryo. The inventive method treats the NP (such as IVF- or NT-derived) embryo at the one-cell or cleavage stage prior to compaction with high osmolality followed by high speed centrifugation. The high osmolality treatment enlarges the periviteline space, and with centrifugation, enables the separation of the lipids from the cytoplasm. The lipid-separated embryos after high osmolality treatment have been successfully cryopreserved and later recovered and transferred to produce live offspring.
Claims5
A method for lipid separation of an IVP porcine embryo comprising
- (a) producing an IVP porcine embryo at the one-cell or cleavage stage prior to compaction,
- (b) exposing such embryo to a medium with a pre-selected osmolality for a short period of time to produce a condensed embryo, and
- (c) centrifuging such condensed embryo at a pre-selected speed and for a pre-selected duration to produce a lipid-separated embryo.
The method of claim 4, wherein said centrifugation condition ranges from about 6 to about 20 minutes at 13,400×g speed.
A method of cryopreservation and recovery of in-vitro-produced porcine embryos comprising:
- (a) producing in-vitro-produced porcine embryos at the one-cell or cleavage stage prior to compaction,
- (b) exposing said embryos to a pre-selected osmolality for a short period of time to produce condensed embryos,
- (c) centrifuging said condensed embryos to produce lipid-separated embryos,
- (d) culturing said lipid-separated embryos to the blastocyst stage to produce lipid-separated blastocysts,
- (e) cryopreserving said blastocysts by vitrification or freezing,
- (f) recovering said blastocysts by warming, rehydrating, and removing the zona pellucida, and
- (g) transferring said zona-removed blastocysts into a recipient.
Dependencies are inferred from the claim text.