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Effect of maternal serum on viability and function of early human placental explants

  • Jahanara Begum-Hasan
  • Mary Senterman
  • Peter Gillett
  • Charlotte Laplante Branchaud
  • Beverley E. Pearson Murphy
Growth, Differentiation And Senescence

Summary

Fetal bovine serum (FBS) is frequently used to supplement chemically defined media such as Ham’s F10 when studying placental explant cultures. However in vitro production of hormones is usually declining by the 2nd or 3rd day and is short-lived (7 to 10 days). In this study we explored the use of human maternal serum (HMS) from early gestation as the medium supplement to Ham’s F10. Early placental hormone production was compared using two concentrations of FBS and HMS. On Day 3 of incubation, progesterone production in 10% HMS was 12-fold increased over that in 10% FBS, estradiol production was increased 10-fold, and βhCG production more than 3-fold. When the serum concentrations were increased to 40%, the results in all cases were similar to those at 10%. Preliminary characterization studies revealed that the stimulatory activity of HMS is heat-labile, neither extractable into organic solvent (diethyl ether) nor dialyzable, suggesting that it is protein in nature. In a long-term incubation, compared with FBS (7 days), HMS permitted survival of culture up to 30 days, judged both histologically and biochemically. We conclude that HMS provides substance(s), probably protein in nature, not present in FBS or non-pregnant human serum, which are important for human placental viability and function in vivo.

Key words

placenta explant culture human maternal serum 

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References

  1. 1.
    Ahmed, N. A. Studies on the regulation of chorionic gonadotropin production in explant cultures of human placenta. Ph.D. Thesis, Department of Physiology, McGill University, Montreal, Canada, 1990.Google Scholar
  2. 2.
    Ahmed, N. A.; Murphy, B. E. P. The effects of various hormones on human chorionic gonadotropin production in early and late placental explant cultures. Am. J. Obstet. Gynecol. 159:1220–1227; 1988.PubMedGoogle Scholar
  3. 3.
    Andrews, R. V.; Phillips, H. J. Instability of metabolic quotients obtained from tissue cultures. Proc. Soc. Exp. Biol. Med. 103:160–163; 1960.PubMedGoogle Scholar
  4. 4.
    Barnes, D.; Sato, G. Methods for growth of cultured cells in serum-free medium. Anal. Biochem. 102:255–270; 1980.PubMedCrossRefGoogle Scholar
  5. 5.
    Begum-Hasan, J.; Murphy, B. E. P. Stimulation of placental progesterone production by 19-nortestosterone and C19 steroids in early human pregnancy. J. Clin. Endocrinol. Metab. 75:838–845; 1992.PubMedCrossRefGoogle Scholar
  6. 6.
    Biggers, J. D.; Rinaldine, L. M.; Webb, M. The study of growth factors in tissue culture. In: Symposia of the Society for Experimental Biology, Sydney; The biological action of growth substances. London, UK: Cambridge University Press; 1957:264–297.Google Scholar
  7. 7.
    Branchaud, C. L.; Goodyer, C. G.; Guyda, H. J., et al. A serum-free system for culturing human placental trophoblast. In Vitro Cell. Dev. Biol. 26:865–870; 1990.PubMedCrossRefGoogle Scholar
  8. 8.
    Branchaud, C. L.; Goodyer, C. G.; Lipowski, L. S. Progesterone and estrogen production by placental monolayer cultures: effect of dehydroepiandrosterone and luteinizing hormone-releasing hormone. J. Clin. Endocrinol. Metab. 56:761–766; 1983.PubMedGoogle Scholar
  9. 9.
    Cobb, J. P.; Walker, D. G. Effect of heterologous, homologous, and autologous serums on human normal and malignant cellsin vitro. JNCI 27:1–15; 1961.PubMedGoogle Scholar
  10. 10.
    Fell, H. B. The development of organ culture. In: Balls, M.; Monnickendan, M., eds. Organ culture in biomedical research. London: Cambridge University Press; 1976:1–13.Google Scholar
  11. 11.
    Hodges, G. M. A review of methodology in organ culture. In: Balls, M.; Monnickendan, M., eds. Organ culture in biomedical research. London: Cambridge University Press; 1976:15–59.Google Scholar
  12. 12.
    Huot, R. I.; Foidart, J. M.; Stromberg, K. Effects of culture conditions on the synthesis of human chorionic gonadotropin by placental organ cultures. In Vitro 15:497–502; 1979.PubMedCrossRefGoogle Scholar
  13. 13.
    Hsu, T. C. Mammalian chromosomesin vitro XIII. Cyclic and directional changes of population structure. JNCI 25:1339–1353; 1960.PubMedGoogle Scholar
  14. 14.
    Klopper, A. Review article: The new placental proteins. Placenta 1:77–89; 1980.PubMedCrossRefGoogle Scholar
  15. 15.
    Marcus, G. J.; Durnford, R. Estradiol assay by microtitre plate enzyme immunoassay. J. Steroid Biochem. 28:207–212; 1988.CrossRefGoogle Scholar
  16. 16.
    Morrish, D. W.; Bhardwaj, D.; Dabbagh, L. K., et al. Epidermal growth factor induces differentiation and secretion of human chorionic gonadotropin and placental lactogen in normal human placenta. J. Clin. Endocrinol. Metab. 65:1282–1290; 1987.PubMedGoogle Scholar
  17. 17.
    Mujumdar, A. S.; Murthy, U.; Das, M. A new trophoblast-derived growth factor from human placenta: purification and receptor identification. Biochemistry 25:627–634; 1986.CrossRefGoogle Scholar
  18. 18.
    Munsick, R. A. Human fetal extremity lengths in the interval from 9 to 21 menstrual weeks of pregnancy. Am. J. Obstet. Gynecol. 149:883–887; 1984.PubMedGoogle Scholar
  19. 19.
    Murphy, B. E. P. Some studies of the protein-binding of steroids and their application to the routine micro and ultramicro measurement of various steroids in body fluids by competitive protein-binding radioassay. J. Clin. Endocrinol. Metab. 27:973–990; 1967.PubMedGoogle Scholar
  20. 20.
    Murphy, W. H.; Bullis, C.; Landau, B. J., et al. Effects of heterologous sera on the modal distribution of variants in four strains of human epithelial cells. Cancer Res. 22:906–917; 1962.PubMedGoogle Scholar
  21. 21.
    Ogino, M.; Kinoshita, K.; Satoh, K., et al. Metabolism of14C-pregnenolone in the placenta throughout pregnancy in organ culture. Endocrinol. Jpn. 30:631–635; 1983.PubMedGoogle Scholar
  22. 22.
    Ogino, M.; Satoh, M.; Katsutuki, K., et al. Viability of the human placental villi in organ culture. Arch. Gynecol. 234:235–246; 1984.PubMedCrossRefGoogle Scholar
  23. 23.
    Paul, S.; Das, C.; Jailkhane, B. L., et al. Progesterone synthesis by human placental syncytiotrophoblastin vitro-preferred precursor and effect of hCG. J. Steroid Biochem. 14:311–313; 1981.PubMedCrossRefGoogle Scholar
  24. 24.
    Riopel, L.; Branchaud, C. L.; Goodyer, C. G., et al. Growth-inhibitory effect of TGF-β on human fetal adrenal cells in primary monolayer culture. J. Cell Physiol. 140:233–238; 1989.PubMedCrossRefGoogle Scholar
  25. 25.
    Siiteri, P. K.; MacDonald, P. C. The utilization of circulating dehydroisoandrosterone sulfate for estrogen synthesis during human pregnancy. Steroids 2:713–730; 1963.CrossRefGoogle Scholar
  26. 26.
    Stromberg, K.; Azizkhan, J. C.; Speeg, K. V. Isolation of functional human trophoblast cells and their partial characterization in primary cell culture. In Vitro 14:631–639; 1978.PubMedCrossRefGoogle Scholar
  27. 27.
    Tan, S. Y.; Murphy, B. E. P. Specificity of the progesterone-binding globulin of the guinea pig. Endocrinology 94:122–127; 1974.PubMedCrossRefGoogle Scholar
  28. 28.
    Veldhuis, J. D.; Azimi, P.; Garmey, J., et al. Biopotential actions of estrogen on progesterone biosynthesis by ovarian cells. II. Relation of estradiol’s stimulatory actions to cholesterol and progesterone metabolism in cultured swine granulosa cells. Endocrinology 117:1076–1083; 1985.PubMedGoogle Scholar
  29. 29.
    Winikoff, J.; Braunstein, G. D. In vitro secretory patterns of human chorionic gonadotrophin, placental lactogen and pregnancy-specific β1-glycoprotein. Placenta 6:417–422; 1985.PubMedCrossRefGoogle Scholar
  30. 30.
    Winkel, C. A.; Snyder, J. M.; MacDonald, P. C., et al. Regulation of cholesterol and progesterone synthesis in human placental cells in culture by serum lipoproteins. Endocrinology 16:1054–1060; 1980.Google Scholar
  31. 31.
    Wu, M. C.; Fischer, R. A. Granulocyte and macrophage colony-stimulating factor from human placental conditioned medium. Biochemistry 19:3846–3850; 1980.PubMedCrossRefGoogle Scholar

Copyright information

© Tissue Culture Association 1993

Authors and Affiliations

  • Jahanara Begum-Hasan
    • 1
  • Mary Senterman
    • 1
  • Peter Gillett
    • 1
  • Charlotte Laplante Branchaud
    • 1
  • Beverley E. Pearson Murphy
    • 1
  1. 1.Reproductive Physiology Unit, Montreal General Hospital, Departments of Medicine, Pathology, and Obstetrics & Gynecology and Centre for the Study of ReproductionMcGill UniversityMontrealCanada

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