Journal of Reproduction & Infertility

Journal of Reproduction & Infertility

Effects of different media on fallopian tube epithelial cells culture and human embryo development

Authors
Maerefat Ghaffari
Mohammad Mehdi Akhondi
Mahnaz Heidari
Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR, Tehran, Iran
Abstract
The advantage of fallopian tube epithelial cell (FTEC) co-culture system on development of human embryo has been shown. However, lack of appropriate media for both FTEC and embryo is one of the disadvantages of this system. The aim of the present study was to determine the effects of different culture media on both FTEC and human embryo development. Healthy fallopian tube samples were removed from 19 previously fertile women under 40 ages undergoing total abdominal hysterectomy. Epithelial cell-rich suspension was prepared mechanically and enzymatically and cultured in three different media (DMEM/F12, RPMI-1640 and Ham’s F10). After four passages, the cells were cultured on plastic to obtain unpolarised confluent cells, or seeded into matrigel to produce polarized cells for 7 days. Viability of cells during and after passages was evaluated by neutral red and trypan blue vital staining. In addition, 117, 45 and 48 surplus 4-8 cell human embryos were cultured in Ham’s F10, RPMI and DMEM/F12 for 120 hours, respectively. Their morphology were assessed daily. Cellular vitality during culture in DMEM/F12 was 100%, which was significantly different from that in RPMI and Ham’s F10 (95% and 93%, respectively)(P < 0.05). However, after cellular proliferation and passages, no significant difference was observed for different media. Moreover, a significantly higher ratio of embryos reached the morula stage in Ham’s F10 (79.4%) than RPMI (68.8%) and DMEM/F112 (62.5%)(P < 0.05). It is concluded that DMEM/F12 was superior to other media during cell proliferation and passage of FTEC. However, after cell passage and during co-culture of embryos with FTEC, Ham’s F10 was a more appropriate medium than the other two.
Keywords
CO
culture
Culture media
Embryo Development
Fallopian tube
Human
Erbach G.T., Lawitts J.A., Papaioannou V.E., Biaggers J.D. Differential growth of the mouse preimplantation embryo in chemically defined media. Biol Reprod. 1994;50(5):1027-33.
Harlow G.M., Quinn P. Development of preimplantation mouse embryos in vivo and in vitro. Aust J Biol Sci. 1982;35(2):187-93.
June T., Fischer B. Correlation between diameter and DNA or protein synthetic activity
Carney E.W., Foote R.H., Effects of super ovulation, embryo recovery, culture system and embryo transfer on development of rabbit embryos in vivo and in vitro. J Reprod Fertil. 1990;89(2):543-51.
Telford N.A., Watson A.J., Schultz G.A. Transition from maternal to embryonic control in early mammalian development: a comparison of several species. Mol Reprod Dev. 1990; 26(1): 90-100. Review.
Braude P., Bolton V., Moore S. Human gene expression first occurs between the four- and eight-cell stages of preimplantation development. Nature. 1988;332(6163):459-61.
FitzGerald L., DiMattina M. An improved medium for long-term culture of human embryos overcomes the in vitro developmental block and increases blastocyst formation. Fertil Steril. 1992; 57(3):641-7.
Thibodeaux J.K., Godke R.A. In vitro enhancement of early-stage embryos with co-culture. Arch Pathol Lab Med. 1992 ;116(4):364-72. Review.
Wiemer K.E., Hoffman D.I., Maxson W.S., Eager S., Muhlberger B., Fiore I., Cuervo M. Embryonic morphology and rate of implantation of human embryos following co-culture on bovine oviductal epithelial cells. Hum Reprod. 1993; 8 (1):97-101.
Menezo Y.J., Guerin J.F., Czyba J.C. Improvement of human early embryo development in vitro by coculture on monolayers of Vero cells. Biol Reprod. 1990;42(2):301-6.
Quinn P., Margalit R. Beneficial effects of co culture with cumulus cells on blastocyst formation in a prospective trial with supernumerary human embryos. J Assist Reprod Genet. 1996;13(1):9-14.
Vlad M., Walker D., Kennedy R.C. Nuclei number in human embryos co-cultured with human ampullary cells. Hum Reprod. 1996; 11 (8):1678-86.
Bongso A., Ng S.C., Fong C.Y., Ratnam S. Cocultures: a new lead in embryo quality improvement for assisted reproduction.Fertil Steril. 1991;56(2):179-91. Review.
Biallie H.S., Pacey A.A., Warren M.A. Greater numbers of human spermatozoa associate with endosalpingeal cells derived from the isthmus compared with those from the ampulla. Hum Reprod.1997;12:1985-1992.
Babich M., Borenfreund E. Application of the neutral red cytotoxicity assay to in vitro toxicology. ATLA 1991;18:129-144.
Wagh P.V., Lippes J. Human oviductal fluid proteins. V. Identification of human oviductin-I as alpha-fetoprotein. Fertil Steril. 1993;59(1):148-56.
Joo B.S., Kim M.K., Na Y.J., Moon H.S., Lee K.S., Kim H.D. The mechanism of action of coculture on embryo development in the mouse model: direct embryo-to-cell contact and the removal of deleterious components. Fertil Steril. 2001;75(1):193-9.
Barmat L.I., Worrilow K.C., Paynton B.V. Growth factor expression by human oviduct and buffalo rat liver coculture cells. Fertil Steril. 1997; 67(4):775-9.
Ghaffari M., et al. Leukaemia inhibitory factor in human postmenopausal endometrial and Fallopian tube cells in vivo and in vitro. 11th world congress on human reproduction, Montreal, Canada (2002)P-012.
Murray C.A., Clarke H.J., Tulandi T., Tan S.L.Inhibitory effect of human ydrosalpingeal fluid on mouse preimplantation embryonic development is significantly reduced by the addition of lactate. Hum Reprod.1997;12(11):2504-7.
Yeung W.S.B., Ho P.C., Lau E.Y.L., Chan S.T.H. Improved development of human embryos in vitro by a human oviducal cell co-culture system. Human reproduction. 1992;7:1144-1149.
Bongso A, Ng SC, Sathananthan A.H., Ng PL, rauff M., Ratnam S.S. Establishment of human ampullary cell cultres. Hum Reprod. 1989;4:486-494.
Kazuhiro Takeuchi, Yukihiro Nagata, Bruce A., Sandow, Gary D, Hodgen. Primary culture of human fallopian tube epithelial cells and co-culture of early mouse per-embryos. Molecular reproduction & development. 1992;32:236-242.
Patek E. The epithelium of the human Fallopian tube. A surface ultrastructural and cytochemical study. Acta Obstet Gynecol Scand. 1974;31(Suppl):1-28.
Donnez J., Casanas-Roux F., Caprasse J., Ferin J., Thomas K. Cyclic changes in ciliation, cell height, and mitotic activity in human tubal epithelium during reproductive life. Fertil Steril. 1985;43(4):554-9.
Ghaffari M. Effects of artificial extracellular matrix on function of human endometrial epithelial cells in vitro. Med J Reprod Infertility. 2000;2:40-49.
Sugrue S.P., Hay E.D. Response of basal epithelial cell surface and Cytoskeleton to solubilized extracellular matrix molecules.J Cell Biol. 1981;91(1):45-54.
Gardner D.K., Lane M. Embryo culture system: In "in vitro fertilization". In Trounson A, Gardner DK (Eds). 1993;PP:85-114.
Baillie H.S., Pacey A.A., Warren M.A., Scud more I.W., Barratt C.L. Greater numbers of human spermatozoa associate with endosalpingeal cells derived from the isthmus compared with those from the ampulla. Hum Reprod. 1997;12(9): 1985-92.
Quinn P., Kerin J.F., Warnes G.M. Improved pregnancy rate in human in vitro fertilization with the use of a medium based on the composition of human tubal fluid. Fertil Steril. 1985;44(4):493-8.
Feng H.L., Wen X.H., Presser S.C. Effect of different co-culture system in early human embryo development. Hum Reprod. 1996;11(7): 1525-8.
Journal of Reproduction & Infertility
Volume 4, Issue 1
Winter 2003
Pages 5-16