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Effect of Vero Cell Coculture on the Development of Frozen–Thawed Two-Cell Mouse Embryos

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Abstract

Purpose: Our purpose was to evaluate the beneficial effects of long-term coculture of Vero cells on the development of frozen–thawed two-cell mouse embryos.

Methods: Two-cell mouse embryos were frozen slowly with 1,2-propandiol and sucrose as cryoprotectants and thawed rapidly, followed by stepwise dilution. Vero cells were cultured in drops of RPMI 1640 to establish monolayers. Frozen–thawed embryos were cultured alone (control) or cocultured with Vero cells. The rate of development in both groups was compared.

Results: After 4 days of culture, significantly more embryos in coculture were developed to expanded blastocysts (61 vs 37% for controls; P ≤ 0.0001). In addition, on the fifth day of cultivation, more embryos in coculture showed the potential of hatching from the zona pellucida (26 vs 7% in controls; P ≤ 0.0001). The rate of degeneration in coculture was also much lower than in controls (6 and 15%, respectively).

Conclusions: Coculture of cryopreserved preimplantation-stage embryos with Vero cells seems to be a useful tool to eliminate the postthaw deleterious effect of freezing and also to obtain better-quality embryos appropriate for transfer.

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REFERENCES

  1. Mahdavan MM, Miller MM: Deleterious effect of equilibration and temperature on the toxicity of propandiol during cryopreservation of mouse zygotes. J Assist Reprod Genet 1997;14:51-54

    Google Scholar 

  2. Massip A, Mermillod P, Vanlangendonckt A, Touze JL, Dessy F: Survival and viability of fresh and frozen-thawed in vitro bovine blastocysts. Reprod Nutr Dev 1995;35:3-10

    Google Scholar 

  3. Van Der Elst J, Van Den Abeel E, Van Streiteghem A: Prospective randomized study on the cryopreservation of human embryos with dimethylsulfoxide or 1, 2-propandiol protocol. Fertil Steril 1995;63:92-100

    Google Scholar 

  4. Shoukir Y, Chardonens D, Campana A, Biscof P, Sakkas D: The role of development and time of transfer play different roles in influencing the viability of human blastocysts. Hum Reprod 1998;13:676-681

    Google Scholar 

  5. Goto K, Iwai N, Takuma Y, Nakanishi Y: Co-culture of in vitro fertilized bovine embryos with different cell monolayers. J Anim Sci 1992;70:1449-1453

    Google Scholar 

  6. Rieger D, Grisart B, Semple E, Van Iangendonck TA, Betteridge KJ, Dessy F: Comparison of the effect of oviductal cell coculture and oviductal cell conditioned medium on the development and metabolic activity of cattle embryos. J Reprod Fertil 1995;105:91-98

    Google Scholar 

  7. Murayama S, Yamano S, Kobayashi T, Ishikawa H, Kunikata K, Anon T: Successful freezing of unfertilized mouse oocytes and effect of coculture in oviducts on development of in vitro fertilized embryos after thawing. J Assist Reprod Genet 1994;11:156-161

    Google Scholar 

  8. Ouhibi N, Hamidi J, Guillad J, Menezo Y: Co-culture of 1-cell mouse embryos on different cell supports. Hum Reprod 1990;5:737-743

    Google Scholar 

  9. Ball BA, Miller PG: Survival of equine embryos co-cultured with equine oviductal epithelium from the four-to eight-cell to blastocyst stage after transfer to synchronous recipient mares. Theriogenology 1992;37:979-991

    Google Scholar 

  10. Watson AJ, Watson PH, Warnes D, Walker SK, Armstrong DT, Seamark RF: Preimplantation development of in vitro-matured and in vitro-fertilized ovine zygotes: Comparison between coculture on oviduct epithelial cell monolayers and culture under low oxygen atmosphere. Biol Reprod 1994;50:715-724

    Google Scholar 

  11. Bongso AM, Ng SC, Sathanthan AH, Ng PL, Rauff M, Ratnam SS: Improved quality of human embryos when cocultured with human ampullary cells. Hum Reprod 1989;4:706-713

    Google Scholar 

  12. Soong YK, Lai YM, Chang SY, Wang ML, Chang MY, Lee CL: A successful pregnancy after subzonal insertion with epididymal sperm and coculture on Vero cell monolayer. Fertil Steril 1993;59:1308-1310

    Google Scholar 

  13. Khalifa EA, Tucker MJ, Hunt PP, Hamidi J: Improved hatching in mouse embryos brought about by combined partial zona dissection and coculture. Hum Reprod 1993;8:599-603

    Google Scholar 

  14. Tucker MJ, Kort HI, Toledo AA, Morton PC, Wright G, Ingargiola PE, Sweitzer CL: Effect of coculture on subsequent survival and implantation of cryopreserved human embryos. J Assist Reprod Genet 1995;12:689-692

    Google Scholar 

  15. Ciriminna R, Palermo R, Cefalu E, et al.: Vero cell Short time coculture improves pregnancy rate after ICSI. J Assist Reprod Genet 1997;14(Suppl):76S

    Google Scholar 

  16. Takagi M, Otoi T, Suzuki T: Survival rate of frozen-thawed bovine IVM/IVF embryos in relation to post-thaw exposure time in two cryoprotectants. Cryobiology 1993;30:466-469

    Google Scholar 

  17. Mahmoudzadeh AR, Van soon A, Bols P, Yesbaert MT, de kruif A: Optimization of a simple vitrification procedure for bovine embryos produced in vitro: Effect of developmental stage, two-step addition of cryoprotectant and sucrose dilution on embryonic survival. J Reprod Fertil 1995;103:33-39

    Google Scholar 

  18. Citadini E, Ciriminna R, Palermo R, Cefalu E, Manno M, Ruvoldo G, Napoli P, Agrifoglio V: Coculture on Vero cells for embryo selection after ICSI or embryo thawing. J Assist Reprod Genet 1997;14(Suppl):PS-06-4

    Google Scholar 

  19. Menezo Y, Guerin JF, Czyba JC: Improvement of human early embryo development in vitro by coculture on monolayers of Vero cells. Biol Reprod 1990;42:301-306

    Google Scholar 

  20. Paria BC, Dey SK: Preimplantation embryo development in vitro cooperative interactions among embryos and role of growth factors. Proc Natl Acad Sci USA 1990;87:4756-4760

    Google Scholar 

  21. Lai YM, Wang HS, Lee CL, Lee JD, Huang HY, Lee JF, Soong YK: Insulin-like growth factor binding-proteins produced by Vero cells, human oviductal cells and human endometrial cells, and the role of insulin-like growth factor binding-protein 3 in mouse co-culture systems. Hum Reprod 1996;11:1281-1286

    Google Scholar 

  22. Maguiness SD, Shrimanker K, Djahanbakhsh O, Grudzinskas JG: Oviduct proteins. Contemp Rev Obstet Gynaecol 1992;4:42-50

    Google Scholar 

  23. Johnson MH, Nasr-Esfanani MH: Radical solutions and cultural problems. BioEssays 1994;16:31-38

    Google Scholar 

  24. Nasr Esfahani MH, Winston NJ, Johnson MH: Effect of glucose, glutamine, EDTA and oxygen tension on the concentration of reactive oxygen species and on development of the mouse preimplantation embryos in vitro. J Reprod Fertil 1992;96:219-231

    Google Scholar 

  25. Lai YM, Lee JD, Wang ML, Chan PJ, Soong YK, Lee LJ, Pao CC: Vero cell coculture improved the development of mouse embryos in the presence of human papillomavirus DNA. J Assist Reprod Genet 1997;14(Suppl):183

    Google Scholar 

  26. Leibo SP, Martino A, Kobayashi S, Pollard JW: Stage dependence sentivity of oocytes and embryos to low temperatures. Anim Reprod Sci 1996;42:45-53

    Google Scholar 

  27. Joly C, Bchini O, Boulekbache H, Testart J, Maro B: Effects of 1, 2-propandiol on the cytoskeletal organization of the mouse oocyte. Hum Reprod 1992;7:374-378

    Google Scholar 

  28. Valojerdi MR, Nematollahi N, Hosseini A, Mozdarani H: The effect of Vero cell on development of one and two cell mouse embryos. MEFSJ 1997;1:35-41

    Google Scholar 

  29. Testart J, Lassalle B, Balaisch-Allart J: High pregnancy rate after early human embryo freezing. Ferti Steril 1986;46:268-272

    Google Scholar 

  30. Veek LL: Freezing of preembryos: Early vs late stages. J Assist Reprod Genet 1993;10:181-185

    Google Scholar 

  31. Jones JM, Trounson AO, Gardner DK, Kausche A, Lolatgis N, Wood C: Evaluation of a culture protocol for successful blastocyst development and pregnancy. Hum Reprod 1998;13:169-177

    Google Scholar 

  32. Gardner DK, Lane M: Culture of viable human blastocysts in defined sequential serum-free media. Hum Reprod 1998;13(Suppl 3): 148-159

    Google Scholar 

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Authors and Affiliations

  1. Department of Anatomy, Kerman University of Medical Sciences, Kerman, Iran

    Nooreddin Nematollahi

  2. Department of Anatomy, Tarbiat Modarres University, Tehran, Iran

    Nooreddin Nematollahi & Mojtaba Rezazadeh Valojerdi

  3. Royan Institute, Jihad Daneshgahi, Medical University of Iran, Tehran, Iran

    Mojtaba Rezazadeh Valojerdi & Mojtaba Rezazadeh Valojerdi

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  1. Nooreddin Nematollahi
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  2. Mojtaba Rezazadeh Valojerdi
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Nematollahi, N., Rezazadeh Valojerdi, M. Effect of Vero Cell Coculture on the Development of Frozen–Thawed Two-Cell Mouse Embryos. J Assist Reprod Genet 16, 380–384 (1999). https://doi.org/10.1023/A:1020598031275

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  • DOI: https://doi.org/10.1023/A:1020598031275

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