Summary
Trophoblast is the major functional cell type of the placenta. The purpose of this study was to devise a means to isolate trophoblast cells from the monkey placenta and to examine their capacity to differentiate in vitro. Methods originally devised for the isolation of human cytotrophoblast cells produced poor yields and a low degree of purity when applied to the near-term rhesus monkey placenta. However, a procedure has been developed which allows the isolation of a cell population consisting of more than 95% cytotrophoblast based on intermediate filament immunocytochemistry. The cells sedimented between densities of 1.040 and 1.053 g/ml on continuous Percoll density gradient centrifugation. When maintained in culture the cells adhered and formed aggregates of mononuclear cells by 24 h. By 5 d in culture, immunofluorescent staining using antidesmoplakin and antinuclear antibodies revealed that most colonies consisted of large multinucleated masses similar to syncytiotrophoblast. These results demonstrate trophoblast cells from monkey placental villi can be isolated with a high degree of purity and undergo morphologic, differentiation in vitro. This preparation should enable investigators to study many functional characteristics of these cells throughout gestation.
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Bax, C. M. R.; Ryder, T. A.; Mobberley, M. A., et al. Ultrastructural changes and immunocytochemical analysis of human placental trophoblast during short-term culture. Placeta 10:170–194; 1989.
Bierings, M. B.; Adriaansen, H. J.; van Dijk, J. P. The appearance, of transferrin receptors on cultured human cytotrophoblast and in vitro-formed syncytiotrophoblast., Placenta 9:387–396; 1988.
Boyd, J. D.; Hamilton, W. J. The human placenta. London: W. Heffer and Sons: 1970:167–174.
Carter, J. E. Morphologic evidence of syncytial formation from the cytotrophoblast cells. Obstet. Gynecol. 23:647–656; 1964.
Daniels-McQueen, S.; Krichevsky, A.; Boime, I. Isolation and characterization of human cytotrophoblast cells. Troph. Res. 2:423–445; 1987.
Douglas, G. C.; King, B. F. Isolation of pure, villous cytotrophoblast from term human placenta using immunomagnetic microspheres. J. Immunol. Methods 119:259–268; 1989.
Enders, A. Formation of syncytium from cytotrophoblast, in the human placenta. Obstet. Gynecol. 25:378–386; 1965.
Feinman, M. A.; Kliman, H. J.; Caltabiano S., et al. 8-Bromo-3′5′-adenosine monophosphate stimulates the endocrine activity of human cytotrophoblasts in culture. J. Clin. Endocrinol. Metab. 63:1211–1217; 1986.
Kao, L. C.; Caltabiano, S.; Wu, S., et al. The human villous cytotrophoblast: interactions with extracellular, matrix proteins, endocrine function, and cytoplasmic, differentiation in the absence of syncytium formation. Dev. Biol. 130:693–702; 1988.
King, N. W. Simian models of acquired immunodeficiency syndrome (AIDS): a review. Vet. Pathol. 23:345–353; 1986.
Kliman, H. J.; Nestler, J. E.; Sermasi., E., et al. Purification, characterization and in vitro differentiation of cytotrophoblasts from human term placentae. Endocrinology 118:1567–1582; 1986.
Lobo, J.; Bellino, F. L..; Siegel, A. Characterization of isolated cells in primary culture from human term placenta by electron microscopy and immunohistochemistry. Troph. Res. 2:447–460; 1987.
Midgley, A. R.; Pierce, G. B.; Deneau, G. A., et al Morphogenesis of syncytiotrophoblast in vivo: an autoradiographic demonstration. Science 141:349–350; 1963.
Moll, R.; Cowin, P.; Kapprell, H-P., et al. Desmosomal, proteins: new markers for identification and classification of tumors. Lab. Invest. 54:4–25; 1986.
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.
Nelson, D. M.; Meister, R. K.; Ortman-Nabi, J., et al. Differentiation and secretory activities of cultured human placental cytotrophoblast. Placenta 7:1–16; 1986.
Pierce, G. B.; Midgley, A. R.; Beals, T. F. An ultrastructural study of differentiation and maturation of trophoblast of the monkey. Lab. Invest. 13:451–463; 1964.
Richart, R. Studies of placental morphogenesis. I. Radioautographic studies of human placenta using tritiated thymidine. Proc. Soc. Exp. Biol. Med. 106:829–831; 1961.
Sopelak, V. M.; Hodgen, G. D. Techniques for research in fetal and neonatal primates. In: Nathanielsz, P. W., ed. Animal models in fetal medicine (III). Ithaca, NY: Perinatology Press; 1984:161–181.
Stromberg, K. The human placenta in cell and organ culture. In: Harris, C. C.; Trump, B. F.; Stoner, G. D., eds. Methods in cell biology, vol. 21B. New York: Academic Press; 1980:227–252.
Stromberg, K.; Huot, R. I. Preferential expression of endogenous type C viral antigen in rhesus, placenta during ontogenesis. Virology 112:365–369; 1981.
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This work was supported by grants HD11658 and RR00169 from the National Institutes of Health, Bethesda, MD.
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Douglas, G.C., King, B.F. Isolation and morphologic differentiation in vitro of villous cytotrophoblast cells from rhesus monkey placenta. In Vitro Cell Dev Biol 26, 754–758 (1990). https://doi.org/10.1007/BF02623616
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DOI: https://doi.org/10.1007/BF02623616