Human Reproduction, Vol. 18, No. 3, 568-571,
March 2003
© 2003 European Society of Human Reproduction and Embryology
A modified cryopreservation method increases the survival of human biopsied cleavage stage embryos
1 Melbourne IVF, 320 Victoria Parade, East Melbourne, Victoria 3002 and 2 Reproductive Services, Royal Women’s Hospital, 132 Grattan Street, Carlton, Victoria 3053, Australia
3 To whom correspondence should be addressed. e-mail: helena{at}mivf.com.au
BACKGROUND: The relatively poor survival rate of human biopsied cleavage stage embryos following cryopreservation is a significant obstacle in the application of preimplantation genetic diagnosis (PGD). We have attempted to improve cryosurvival of biopsied embryos by modifying the standard embryo cryopreservation technique. METHODS: Biopsied embryos were cryopreserved in 1.5 mol/l 1,2-propanediol in the presence of an elevated concentration of sucrose (0.2 mol/l) and human serum albumin was replaced by maternal serum (20% vol:vol). An additional initial thawing step in the presence of 0.3 mol/l sucrose was also included. RESULTS: The proportion of biopsied embryos which survived cryopreservation with 
50% of their blastomeres intact was significantly higher using the modified method (138/185; 75%) than that observed using the standard propanediol method (20/46; 43%; P = 0.022). Total blastomere survival was also significantly increased as a result of the modifications (1010/1513; 67% versus 177/385; 46%; P < 0.001). Six fetal hearts have been detected to date following replacement of biopsied embryos cryopreserved with the modified method. CONCLUSIONS: Survival of human biopsied cleavage stage embryos can be restored to a level similar to that of non-biopsied controls by modification of the cryopreservation procedure. Embryos which have been cryopreserved using the modified method can implant following replacement in utero.
Key words: cryopreservation/embryo/implantation/PGD
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