Cryoloop vitrification yields superior survival of Rhesus monkey blastocysts

BACKGROUND: Vitrification using the cryoloop procedure was evaluated for preservation of non-human primate blastocysts by comparing survival results from two different cryoprotectant mixtures with prior results from controlled rate cooling. METHODS: Rhesus monkey blastocysts were produced by intracy...

Full description

Saved in:
Bibliographic Details
Published in:Human reproduction (Oxford) 2001-09, Vol.16 (9), p.1965-1969
Main Authors: Yeoman, R.R., Gerami-Naini, B., Mitalipov, S., Nusser, K.D., Widmann-Browning, A.A., Wolf, D.P.
Format: Article
Language:English
Subjects:
Animals
Biological and medical sciences
Birth control
Blastocyst - physiology
blastocysts
Cells, Cultured
cryoloop
cryopreservation
Cryopreservation - methods
Cryoprotective Agents
Drug Combinations
Embryo Transfer
Ethylene Glycol
Female
Glycerol
Gynecology. Andrology. Obstetrics
Macaca mulatta
Medical sciences
monkey
Pregnancy
Pregnancy Rate
Rats
Rats, Inbred BUF
Solutions
Sterility. Assisted procreation
vitrification
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:BACKGROUND: Vitrification using the cryoloop procedure was evaluated for preservation of non-human primate blastocysts by comparing survival results from two different cryoprotectant mixtures with prior results from controlled rate cooling. METHODS: Rhesus monkey blastocysts were produced by intracytoplasmic sperm injection of mature oocytes from cycling females stimulated with recombinant human hormones. Morphologically well-formed blastocysts were divided between Procedure A (2.8 mol/l dimethylsulphoxide and 3.6 mol/l ethylene glycol with 0.65 mol/l sucrose and 25 μmol/l Ficoll in TALP-HEPES with 20% fetal bovine serum (TH20)) and Procedure B (3.4 mol/l glycerol and 4.5 mol/l ethylene glycol in TH20). After >48 h in liquid nitrogen, the removal of cryoprotectants was accomplished in the presence of a 3-step series of decreasing sucrose concentrations in TH20. Surviving embryos were co-cultured on buffalo rat liver cells. RESULTS: Of 16 blastocysts vitrified via Procedure A, 38% survived with minimal lysis and only one hatched in culture; in contrast, of 33 blastocysts vitrified by Procedure B, 85% survived and 71% hatched. Of 22 blastocysts cryopreserved by conventional slow cooling, 36% survived and 6% hatched. Transfer into three recipients, each with two embryos vitrified with Procedure B, resulted in a successful twin-term pregnancy. CONCLUSION: Modified cryoloop vitrification with a final solution of 3.4 mol/l glycerol and 4.5 mol/l ethylene glycol is a promising procedure for preserving Rhesus monkey blastocysts that is simple, rapid, and inexpensive.
ISSN:0268-1161
1460-2350
DOI:10.1093/humrep/16.9.1965