Skip to main content
Log in

Assessment of hamster blastocysts derived from eight-cell embryos cultured in hamster embryo culture medium-2 (HECM-2): Cell numbers and viability following embryo transfer

  • Special Contributions
  • Published:
Journal of in Vitro Fertilization and Embryo Transfer Aims and scope Submit manuscript

Abstract

The viability of hamster blastocysts, cultured from the eight-cell stage using hamster embryo culture medium-2, was examined by embryo transfer. Approximately 15–20 cultured hamster blastocysts were surgically transferred unilaterally to uterine horns of pseudopregnant recipient hamsters that had been mated to vasectomized males 3 days previously. Control recipients received in vivo developed, freshly recovered eight-cell embryos or blastocysts on day 2 or 3 of pseudopregnancy, respectively. Of the successful embryo transfers, the experimental group (receiving cultured blastocysts;n=10) gave 51.8% implantations and 28.2% live pups. These values were closely similar to those of the controls; the percentages of control implantations and offspring were 51.1 and 34.0%, respectively, for eight-cell embryo transfer (n=7,P>0.69) and 48.5 and 28.9% for blastocyst transfer (n=6,P>0.52). To evaluate the quality of cultured hamster blastocysts, the following two parameters were examined. (1) The mean number of cells per blastocysts was 24.4±0.7 for cultured blastocysts. This value was similar to that (range, 14–24) obtained in this laboratory for in vivo developed freshly recovered blastocysts. (2) Oxygen consumption analysis revealed that cultured blastocysts actively respired at a level close to that observed with freshly recovered eight-cell embryos (slopes of oxygrams: 0.25 and 0.26, respectively). From these results, it is concluded that hamster blastocysts, cultured from the eight-cell stage, are (a) qualitatively similar to freshly recovered in vivo developed blastocysts and (b) biologically viable as revealed by the production of live offspring upon embryo transfer. Therefore, hamster embryo culture medium-2 could be useful for studies relating to embryo-derived proteins involved in the process of implantation and for the production of transgenic hamster embryos, for example, using embryonic stem cells.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

  1. Iritani A: Current status of biotechnological studies in mammalian reproduction. Fertil Steril 1988;50:543–551

    PubMed  Google Scholar 

  2. Jones GS: Update on in vitro fertilization. Endocrine Rev 1984;5:62–75

    Google Scholar 

  3. Wood C, McMaster R, Rennie G, Trounson A, Leeton J: Factors influencing pregnancy rates following in vitro fertilization and embryo transfer. Fertil Steril 1985;43:245–250

    PubMed  Google Scholar 

  4. George MA, Doe BG: The influence of handling procedures during mouse oocyte and embryo recovery on viability and subsequent development in vitro. J Vitro Fert Embryo Transfer 1989;6:69–72

    Google Scholar 

  5. George AG, Braude PR, Johnson MH, Sweetnam DG: Quality control in the IVF laboratory: In vitro and in vivo development of mouse embryos is unaffected by the quality of water used in culture media. Hum Reprod 1989;4:826–831

    PubMed  Google Scholar 

  6. Bavister BD: Studies on the developmental blocks in cultured hamster embryos.In The Mammalian Preimplantation Embryo: Regulation of Growth and Differentiation in Vitro, BD Bavister (ed). New York, Plenum Press, 1987, pp 219–250

    Google Scholar 

  7. Bavister BD: Regulation of hamster preimplantation embryo development in vitro by glucose and phosphate.In Early Embryo Development and Paracrine Relationships, S Heyner, L Wiley (eds). New York, Alan R. Liss, 1990, pp 79–96

    Google Scholar 

  8. Schini SA, Bavister BD: Two-cell block to development of cultured hamster embryos is caused by phosphate and glucose. Biol Reprod 198839:1183–1192

    PubMed  Google Scholar 

  9. Seshagiri PB, Bavister BD: Glucose inhibits development of hamster 8-cell embryos in vitro. Biol Reprod 1989;40:599–606

    PubMed  Google Scholar 

  10. Seshagiri PB, Bavister BD: Phosphate is required for glucose inhibition of development of hamster 8-cell embryos in vitro. Biol Reprod 1989;40:607–614

    PubMed  Google Scholar 

  11. Monis H, Bavister BD: Development of 4-cell hamster embryos to the blastocyst stage in vitro and its regulation by components of the culture milieu. Reprod Fertil Dev 1990;2:1–9

    PubMed  Google Scholar 

  12. Schini SA, Bavister BD: Normal offspring produced after transfer of hamster embryos grown from 2- to 8-cells in vitro using a chemically defined culture medium. Theriogenol 1990;33:1255–1262

    Google Scholar 

  13. Bavister BD, Leibfried ML, Lieberman G: Development of preimplantation embryos of the golden hamster in a defined culture medium. Biol Reprod 1983;28:235–247

    PubMed  Google Scholar 

  14. Seshagiri PB, Bavister BD: Relative developmental abilities of hamster 2- and 8-cell embryos cultured in HECM-1 and-2. J Exp Zool (accepted)

  15. Orsini MW: The external vaginal phenomena characterizing the stages of the estrous cycle, pregnancy, pseudopregnancy, lactation and the anestrous hamster. Proc Anim Care Panel 1964;11:193–206

    Google Scholar 

  16. Bavister BD: A consistently successful procedure for in vitro fertilization of golden hamster eggs. Gamete Res 1989; 23:139–158

    PubMed  Google Scholar 

  17. Pursel VG, Wall RJ, Rexroad CE Jr, Hammer RE, Brinster RL. A rapid whole-mount staining procedure for nuclei of mammalian embryos. Theriogenol 1985;24:687–691

    Google Scholar 

  18. Snedecor GW, Cochran WG: Statistical Methods. Ames, Iowa State University Press, 1982, pp 290–291

    Google Scholar 

  19. Caro CM, Trounson A: The effect of protein on preimplantation mouse embryo development in vitro. J Vitro Fert Embryo Transfer 1984;1:183–187

    Google Scholar 

  20. Vanderhyden BC, Armstrong DT: Decreased embryonic survival of in vitro fertilized oocytes in rats is due to retardation of preimplantation development. J Reprod Fert 1988;83:851–857

    Google Scholar 

  21. Pope WF. Uterine asynchrony: A cause of embryonic loss. Biol Reprod 1988;39:999–1003

    PubMed  Google Scholar 

  22. Rexroad CE, Powell AM: Asynchronous transfer improves development of sheep embryos co-cultured in serum-free medium. Biol Reprod 1989 40(Suppl 1): 137 (No. 268)

    Google Scholar 

  23. Chang MC: Development and fate of transferred rabbit ova or blastocyst in relation to the ovulation time of recipients. J Exp Zool 1950;114:197–226

    Google Scholar 

  24. Rowson LEA, Moore RM, Lawson RAS: Fertillity following egg transfer in the cow: Effect of method, medium and synchronization of oestrus. J Reprod Fert 1969;18:517–523

    Google Scholar 

  25. Overstrom E: In vitro assessment of blastocyst differentiation.In The Mammalian Preimplantation Embryo: Regulation of Growth and Differentiation in Vitro, BD Bavister (ed). New York, Plenum Press, 1987, pp 95–116

    Google Scholar 

  26. Bavister BD, Andrews JC: A rapid sperm motility bioassay procedure for quality-control testing of water and culture media. J Vitro Fert Embryo Transfer 1988;5:67–75

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Seshagiri, P.B., Bavister, B.D. Assessment of hamster blastocysts derived from eight-cell embryos cultured in hamster embryo culture medium-2 (HECM-2): Cell numbers and viability following embryo transfer. J Assist Reprod Genet 7, 229–235 (1990). https://doi.org/10.1007/BF01129524

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF01129524

Key Words

Navigation