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Apr 25, 2017
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Jun 1, 2005
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Abstract

Successful oocyte fertilization and normal embryonic development of mice were considered the most important diagnostic criteria for the safety of materials and tools used for human in vitro fertilization and embryo transfer (IVF-ET). Therefore, we studied the influence of cumulus cells co-culture and protein supplement within culture medium on percentages of in vitro fertilization (IVF) and normal development of early stages of mouse embryo later. Oocytes were collected and treated with hyaluronidase to remove cumulus cells. Oocytes were divided into four groups namely: Group-1: Oocytes incubated within modified Earl’s medium (MEM) supplied with 10% inactivated bovine amniotic fluid as a protein source and cumulus cells; Group-2: Oocytes incubated with MEM supplied with cumulus cells only; Group-3: Oocytes incubated with MEM supplied with 10% inactivated bovine amniotic fluid only; and Group-4: Oocytes  incubated with MEM free of both protein source and cumulus cells. For IVF, 5-6 oocytes were incubated with active spermatozoa under paraffin oil for 18-20 hours at 37° oC in 5% CO2. Percentages of IVF and embryonic development were then recorded. Best results for IVF and normal embryonic development were achieved from oocytes of Group-1 when compared to the other groups. As compared to Group-1, the percentage of IVF for Group-2 and Group-3 were decreased insignificantly and significantly (P<0.002), respectively. Significant (P<0.01) reduction in the percentages of IVF and normal embryonic development were reported in Group-4 as compared to Group-1. Therefore, it was concluded that the presence of cumulus cells co-culture and bovine amniotic fluid as a protein source within culture medium may have an important role on the fertilizing capacity of spermatozoa and oocytes and normal development of pre-implanted mouse embryo later.

 

 

Keywords

Mouse fertilization embryo culture medium IVF.

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References

  1. ARICI, A., BEHRMAN, H.R. and KEEFE, D.L. 1999. Prostaglandins and prostaglandin-like products in reproduction: Eicosanoids, peroxides and oxygen radicals. In: Reproductive Endocrinology. Yen, S.S.C., Jaffe, R.B., Barbiere, R.L. (eds.). W.B. Saunders Company, Pennsylvania, USA. 4th Edition. Pp: 134-152.
  2. BAE, I.H., FOOTE and R.H. 1980. Maturation of rabbit follicular oocytes in a defined medium of varied osmolality. J. Reprod. Fertil. 59: 11-13.
  3. BARBEHENN, E.K., WALES, R.G. and LOWRY, O.H. 1974. The explanation for the blockade of glycolysis in early mouse embryos. Proc. Natl. Acad. Sci. USA. 71: 1056-1060.
  4. BAVISTER, B.D. 1995. Culture of preimplantation embryos: facts and artifacts. Hum. Reprod. Update. 1: 91-148.
  5. BORNSLAEGER, E.A. and SCHULTZ, R.M. 1985. Regulation of mouse oocyte maturation: Effect of elevating cumulus cells cAMP on oocyte cAMP levels. Biol. Reprod. 33: 698-704.
  6. BUEHR, M. 1995. Embryo technology in laboratory animals. Scand. J. Lab. Anim. Sci. 22: 75-85.
  7. CALVO, L., VANTMAN, D., BANKS, S.M., TEZON, J., KOUKOULIS, G.N., DENNISON, L. and SHERINS, R.J. 1989. Follicular fluid-induced acrosome reaction distinguishes a subgroup of men with unexplained infertility not identified by semen analysis. Fertil. Steril. 52: 1048-1054.
  8. CANIPARI, R. 2000. Oocyte-granulosa cell interactions. Hum. Reprod. Update. 6: 279-289.
  9. CONAGHAN, J., HARDY, K., LEESE, H.J., WINSTON, R.M.L. and HANDYSIDE, A.H. 1998. Culture of human preimplantation embryos to the blastocyst stage: a comparison of three media. Int. J. Dev. Biol. 42: 885-893.
  10. DIEDRICH, K., VAN DER VEN, H., AL-HASANI, S. and KREBS, D. 1988. Ovarian stimulation for in vitro fertilization. Hum. Reprod. 3: 39-44.
  11. DURAN, D.H. 2000. Technical aspect of in vitro embryo production. Internet file, 1-10.
  12. EMERY, B., MILLER, R. and CARRELL, D.T. 2001. Hamster oocyte membrane potential and ion permeability vary with preantral cumulus cell attachment and developmental stage. BMC Develop. Biol. 1: 14-22.
  13. FAKHRILDIN, M-B. M-R. 2000. Materials and Methods. In: The impact of seminal fluid infections and treatment on sperm functions and fertilizing capacity in infertile patients. Ph. D. Thesis. College of Science, Al-Mustansiriya. Pp: 72-84.
  14. FINDLAY, A.L.R. 1984. The fetus and neonate. In: Reproduction and the fetus. Hardy, R.N., Hobsley, M., Saunders, K.B., Fitzsimons, J.T. (eds.). Edward Arnold (Publishers) Ltd. London, UK. pp: 96-129.
  15. FUNAHASHI, H., CANTLY, T.C. and DAY, B.N. 1997. Synchronization of meiosis in porcine oocytes by exposure to dibutyryl cAMP improves developmental competence following in vitro fertilization. Biol. Reprod. 57: 49-53.
  16. GRAAFF, V.D. 2002. Extraembryonic membranes. Development. 6th Edition. Chapter 22; pp: 754-767 (250lec4.html).
  17. HARTSHORNE, G.M. 1989. Steroid production by the cumulus: relationship to fertilization in vitro. Hum. Reprod. 4: 742-745.
  18. IVF.HTM; Internet file. In vitro fertilization: An overview for high school students. P: 1-8.
  19. JOHNSON, M. and EVERITT, B. 1988. The fetus and its preparations for birth. In: Essential Reproduction. Johnson, M., Everitt, B., (eds.). 3rd Edition. Blackwell Scientific Publications. Oxford, UK. Pp: 265-293.
  20. KANE, M.T. and HEADON, D.R. 1980. The role of commercial bovine serum albumin preparations in the culture of one-cell rabbit embryos to blastocysts. J. Reprod. Fertil. 60: 469-475.
  21. KOL, S. and ADASHI, E.Y. 1995. Intraovarian factors regulating ovarian function. Curr. Opin. Obstet. Gynecol. 7: 209-213.
  22. LANE, M. and GARDNER, G.K. 1997. Differential regulation of mouse embryo development and viability by amino acids. J. Reprod. Fert. 109: 153-164.
  23. LANGMAN, J. 1981. Fetal membranes and placenta. In: Medical embryology. Langman, J., (ed.). Williams and Wilkins Company, Baltimore, USA. 4th Edition. Pp: 83-101.
  24. LI, X., MORRIS, L.A-H. and ALLEN, W.R. 2001. Influence of co-culture during maturation on the developmental potential of equine oocytes fertilized by intracytoplasmic sperm injection. Reprod. 121: 925-932.
  25. MADAN, M.L., CHAUHAN, M.S., SINGLA, S.K. and MANIK, R.S. 1994. Pregnancies established from water buffalo (Bubalus bubalis) blastocysts derived from in vitro matured, in vitro fertilized oocytes and co-cultured with cumulus and oviductal cells. Theriogenology 42: 591-600.
  26. MAEDA, J., KOTSUJI, F., NEGAMI, A., KAMITANI, N. and TOMINAGA, T. 1996. In vitro development of bovine embryos in conditioned media from bovine granulosa cells and vero cells cultured in exogenous protein- and amino acid-free chemically defined human tubal fluid medium. Biol. Reprod. 54: 930-936.
  27. MAHADEVAN, M., MILLER, M.M. and MOUTOS, D.M. 1997. Absence of glucose decreases human fertilization and sperm movement characteristics in vitro. Hum. Reprod. 12: 119-123.
  28. MINAMI, N., KATO, H., INOUE, Y., YAMADA, M., UTSUMI, K. and IRITANI, A. 1994. Non species specific effect of mouse oviducts on the development of bovine IVM/IVF embryos by a serum free co-culture. Theriogenology 41: 1435-1445.
  29. MOORE, K.L. 1988. Placenta and fetal membranes. In: Essentials of human embryology. Moore, K.L., (ed.). B. C. Decker Inc., Ontario, Canada. pp: 43-52.
  30. MORI, T., AMANO, T. and SHIMIZU, H. 2000. Roles of gap junctional communication of cumulus cells in cytoplasmic maturation of porcine oocytes cultured in vitro. Biol. Reprod. 62: 913-919.
  31. MOTLIK, J. and FULKA, J. 1981. Fertilization of rabbit oocytes co-cultured with granulos cells. J. Reprod. Fertil. 63: 425-429.
  32. OSHEROFF, J., VISCONTI, P., VALENZUELA, J.P., TRAVIS A.J., ALVAREZ, J. and KOPF, G.S. 1999. Regulation of human sperm capacitation by a cholesterol efflux-stimulated signal transduction pathway leading to protein kinase A-mediated up-regulation of protein tyrosine phosphorylation. Mol. Hum. Reprod. 5: 1017-1026.
  33. PETER, W. and PEARSON, A.E.G. 1971. The laboratory animals: principles and practice. Academic Press. New York, USA. Pp: 226-234.
  34. PIKE, I.L. and ALIKANI, M. 1990. Time-dependent loss of developmental potential when two-celled mouse embryos were retained in culture in excised oviducts. Ann. New York Acad. Scien. 419-423.
  35. SHIMADA, M., MAEDA, T. and TERADA, T. 2001. Dynamic changes of connexin-43, gap junctional protein, in outer layers of cumulus cells are regulated by PKC and PI 3-kinase during meiotic resumption in porcine oocytes. Biol. Reprod. 64: 1255-1263.
  36. SHIMADA, M. and TERADA, T. 2002. FSH and LH induce progesterone production and progesterone receptor synthesis in cumulus cells, a requirement for meiotic resumption in porcine oocytes. Mol. Hum. Reprod. 8: 612-618.
  37. STEPTOE, P.C. and EDWARDS, R.G. 1978. Birth after the re-implantation of a human embryo. Lancet 2: 336-341.
  38. SU, Y-Q., WIGGLESWORTH, K., PENDOLA, F.L., O’BRIEN, M.J. and EPPIG, J.J. 2002. Mitogen-activated protein kinase activity in cumulus cells is essential for gonadotropin-induced oocyte meiotic resumption and cumulus expansion in the mouse. Endocrinology 143: 2221-2232.
  39. TOTEY, S.M., PAWSHE, C.H. and SINGH, G.P. 1993. In vitro maturation and fertilization of buffalo oocytes (Bubalus bubalis): Effect of media hormone and sera. Theriogenology 39: 1135-1171.
  40. VONGPRALUB, T. and KOYANAGI, F. 1994. Effects of oxytocin and granulose cell co-culture on the development of mouse one-cell stage embryos. J. Mamm. Ova. Res. 11: 175-181.
  41. WATSON, A.J., WATSON, P.H., WARNES, D., WALKER, S.K., ARMSTRONG, D.T. and SEAMARK, R.F. 1994. Preimplantation development of in vitro matured and in vitro fertilized ovine zygotes. Comparison between co-culture on oviduct epithelial cell monolayer and culture under low oxygen atmosphere. Biol. Reprod. 50: 715-724.
  42. WEISS, T. and ECKERT, A. 1989. Cotinine levels in follicular fluid and serum of IVF patients: effect on granulose-luteal cell function in vitro. Hum. Reprod. 4: 482-485.
  43. XU, K.P., YADAF, B.R., RORIE, R.W., PLANTE, L., BETTERIDGE, K.J. and KING, W.A. 1992. Development and viability of bovine embryos ferived from oocytes matured and fertilized in vitro and co-culture with bovine oviductal epithelial cells. J. Reprod. Fertil. 94: 33-43.
  44. YADAV, P.S., SAINI, A., KUMAR, A. and JAIN, G.C. 1998. Effect of oviductal cell co-culture on cleavage and development of goat IVF embryos. Animal Reprod. Science 51: 301-306.
  45. YEN, S.S.C. and JAFFE, R.P. 1999. Prolactin in human reproduction. In: Reproductive Endocrinology. Yen, S.S.C., Jaffe, R.B., Barbiere, R.L., (eds.). W.B. Saunders Company, Pennsylvania, USA. 4th Edition. Pp: 257-283.
  46. YEUNG, W.S.B. and NG, E.H.Y. 2000. Laboratory aspects of assisted reproduction. HKMJ. 6: 163-168.
  47. ZHU, J., BARRAT, C.L.R., LIPPES, J., PACEY, A.A., LENTON, E.A. and COOKE, I.D. 1994. Human oviductal fluid prolongs sperm survival. Fertil. Steril. 61: 360-366.