Semin Reprod Med 2002; 20(1): 037-044
DOI: 10.1055/s-2002-23518
Copyright © 2002 by Thieme Medical Publishers, Inc., 333 Seventh Avenue, New York, NY 10001, USA. Tel.: +1(212) 584-4662

Cryopreservation, Thawing, and Transfer of Human Embryos

William Byrd
  • Departments of Obstetrics and Gynecology and Urology, University of Texas Southwestern Medical Center, Dallas, Texas
Further Information

Publication History

Publication Date:
02 April 2002 (online)

ABSTRACT

Cryopreservation of human embryos is a routine procedure in most assisted reproductive technology laboratories. Embryos can be successfully frozen at the pronuclear, multicellular (four to eight blastomeres), and blastocyst stages. Each laboratory has to determine which freezing protocol is best suited for its patients. Freezing protocols have been developed that utilize cryoprotective agents such as glycerol, propanediol, dimethylsulfoxide, and sucrose. The purpose of these cryoprotectants is to help reduce the formation of intracellular ice when using a slow cooling protocol to freeze the embryos. The two most common ways to freeze embryos utilize either low concentrations of cryoprotectants and a slow stepwise freeze or high concentrations of cryoprotectants and a rapid freeze. Frozen-thawed embryos can be transferred to naturally cycling women or to women who have been primed with hormone replacement treatment with equal success.

REFERENCES

  • 1 Trounson A, Mohr L. Human pregnancy following cryopreservation, thawing and transfer of an eight-cell embryo.  Nature . 1983;  305 707-709
  • 2 Assisted Reproductive Technology Success Rates. Atlanta: Centers for Disease Control . 2000
  • 3 Whittingham D G, Leibo S P, Mazur P. Survival of mouse embryos frozen to -196° and -269°C.  Science . 1972;  178 411-414
  • 4 Wilmut I. The effect of cooling rate, warming rate, cryoprotective agent and stage of development on survival of mouse embryos during freezing and thawing.  Life Sci . 1972;  11 1071-1079
  • 5 Cohen J, Simmons R F, Edwards R G, Fehilly C B, Fishel S B. Pregnancies following the storage of expanding human blastocysts.  J In Vitro Fert Embryo Trans . 1985;  2 59-64
  • 6 Fehilly C B, Cohen J, Simons R F, Fishel S B, Edwards R G. Cryopreservation of cleaving embryos and expanding blastocysts in the human: a comparative study.  Fertil Steril . 1985;  44 638-644
  • 7 Lassalle B, Testart J, Renard J-P. Human embryo features that influence the success of cryopreservation with the use of 1,2 propanediol.  Fertil Steril . 1985;  44 645-651
  • 8 Reubinoff B E, Pera V F, Vajta G, Trounson A O. Effective cryopreservation of human embryonic stem cells by open pulled straw vitrification method.  Hum Reprod . 2001;  16 2187-2194
  • 9 Landa V, Tepla O. Cryopreservation of mouse 8-cell embryos in microdrops.  Folia Biol (Praha) . 1990;  36 153-158
  • 10 Leibo S P. Procedures to cryopreserve zygotes and embryos.  Hands-on Cryobiology Course. Indianapolis: American Fertility Society 1993: 69-78
  • 11 Glenister P H, Whittingham D G, Lyon M F. Further studies on the effect of radiation during the storage of frozen 8-cell mouse embryos at -196°C.  J Reprod Fertil . 1984;  70 229-234
  • 12 Becker D, Sevilla M D. The chemical consequences of radiation-damage to DNA.  Adv Rad Biol . 1993;  17 121-180
  • 13 Tedder R S, Zukerman M A, Goldstone A H. Hepatitis B transmission from contaminated cryopreservation tank.  Lancet . 1995;  346 137-140
  • 14 Troup S A, Matson P L, Critchlow J D, Morroll D R, Lieberman B A, Burslem R W. Cryopreservation of human embryos at the pronucleate, early cleavage or expanded blastocyst stages.  Eur J Obstet Gyncol Reprod Biol . 1991;  38 133-139
  • 15 Veeck L L, Amundson C H, Brothman L J. Significantly enhanced pregnancy rates per cycle through cryopreservation and thaw of pronuclear stage oocytes.  Fertil Steril . 1993;  59 1202-1207
  • 16 Quinn P. Cryopreservation of embryos and oocytes. In: Keye WR Jr, Chang RJ, Rebar RW, Soules MR, eds. Infertility, Evaluation and Treatment New York: WB Saunders 1995: 821-840
  • 17 Lelaidier C, de Ziegler D, Freitas S, Olivennes F, Hazout A, Frydman R. Endometrium preparation with exogenous estradiol and progesterone for the transfer of cryopreserved blastocysts.  Fertil Steril . 1995;  63 919-921
  • 18 Horne G, Critchlow J D, Newman M C, Edozien L, Matson P L, Liberman B A. A prospective evaluation of cryopreservation strategies in a two-embryo transfer programme.  Hum Reprod . 1997;  12 542-547
  • 19 Kattera S, Shrivastav P, Craft I. Comparison of pregnancy outcomes of pronuclear and multicellular stage frozen thawed embryo transfers.  J Assist Reprod Genet . 1999;  16 358-361
  • 20 Senn A, Vozzi C, Chanson A, De Grandi P, Germond M. Prospective randomized study of two cryopreservation policies avoiding embryo selection: the pronucleate stage leads to a higher cumulative delivery rate than the early cleavage stage.  Fertil Steril . 2000;  74 946-952
  • 21 Rijnders P M, Jansen C A. The predictive value of day 3 embryo morphology regarding blastocyst formation pregnancy and implantation rate after day 5 transfer following in-vitro fertilization of intracytoplasmic sperm injection.  Hum Reprod . 1998;  13 2869-2873
  • 22 Graham J, Han T, Porter R, Levy M, Stillman R, Tucker M J. Day 3 morphology is a poor predictor of blastocyst quality in extended culture.  Fertil Steril . 2000;  74 495-497
  • 23 Fehilly C B, Cohen J, Simons R F, Fishel S B, Edwards R G. Cryopreservation of cleaving embryos and expanded blastocysts in the human: a comparative study.  Fertil Steril . 1984;  44 638-644
  • 24 Gardner D K, Lane M. Embryo culture systems In: Trounson A, Gardner DK, eds. Handbook of In Vitro Fertilization. New York: CRC Press; 2000: 85-114
  • 25 Van den Abbeel E, Van Steirteghem A. Zona pellucida damage to human embryos after cryopreservation and the consequences for their blastomeres survival and in-vitro viability.  Human Reprod   2000;  15 373-378
  • 26 Magli M C, Gianaroli L, Fortini D, Ferraretti A P, Munne S. Impact of blastomeres biopsy and cryopreservation techniques on human embryo viability.  Hum Reprod . 1999;  14 770-773
  • 27 Joris H, Van den Abbell E, Vos A D, Van Steirteghem A. Reduced survival after human embryo biopsy and subsequent cryopreservation.  Hum Reprod . 1999;  14 2833-2837
  • 28 Kondo I, Suganuma N, Ando T, Furuhashi M, Tomoda Y. Clinical factors for successful cryopreserved-thawed embryo transfer.  J Assist Reprod Genet . 1996;  13 201-206
  • 29 Van der Elst J, van den Abbeel E, Vitrier S, Camus M, Devroey P, van Steirteghem C A. Selective transfer of cryopreserved embryos with further cleavage after thawing increases delivery and implantation rates.  Hum Reprod . 1997;  12 1513-1521
  • 30 Edgar D H, Bourne H, Jericho H, McBain J C. The developmental potential of cryopreserved human embryos.  Mol Cell Endocrinol . 2000;  169 69-72
  • 31 Shoukir Y, Chardonnens D, Campana A, Bischof P, Sakkas D. The rate of development and time of transfer play different roles in influencing the viability of human blastocysts.  Hum Reprod . 1998;  13 676-681
  • 32 Nakayama T, Goto Y, Kanzaki H. Developmental potential of frozen-thawed human blastocysts.  J Asst Reprod Gen . 1995;  12 239-243
  • 33 Tucker M J, Cohen J, Massey J B, Mayer M P, Wiker S R, Wright G. Partial dissection of the zona pellucida of frozen-thawed human embryos may enhance blastocyst hatching, implantation, and pregnancy rates.  Am J Obstet Gynecol . 1991;  165 341-344
  • 34 Check J H, Hoover L, Nazari A, O'Shaughnessy A, Summers D. The effect of assisted hatching on pregnancy rates after frozen embryo transfer.  Fertil Steril . 1996;  65 254-257
  • 35 Mantoudis E, Podsiadly B T, Gorgy A, Venkat G, Craft I L. A comparison between quarter, partial and total laser assisted hatching in selected infertility patients.  Hum Reprod . 2000;  16 2182-2186
  • 36 Schmidt C L, de Ziegler D, Gagliardi C L. Transfer of cryopreserved-thawed embryos: the natural cycle versus controlled preparation of the endometrium with gonadotropin-releasing hormone agonist and exogenous estradiol and progesterone (GEEP).  Fertil Steril . 1989;  52 609-616
  • 37 Sathanandan M, MacNamee M C, Rainsbury P, Wick K, Brendan P, Edwards R G. Replacement of frozen-thawed embryos in artificial and natural cycles: a prospective semi-randomized study.  Hum Reprod . 1991;  6 685-687
  • 38 Muasher S J, Kruithoff C, Simometti S, Oehningr S, Acosta A A, Jones G S. Controlled preparation of the endometrium with exogenous steroids for the transfer of frozen-thawed pre-embryos in patients with anovulatory or irregular cycles.  Hum Reprod . 1991;  6 443-445
  • 39 Yee B, Lin Y P, Chacon R R, Soubra S, Rosen G F, Cassidenti D L. A simplified method of timing frozen embryo transfers.  Am J Obstet Gynecol . 1995;  172 1844-1848
    >