Characterization and differentiation of human first trimester placenta trophoblastic cells in culture

Abstract

Summary ― A preparation of highly enriched isolated cytotrophoblasts was obtained from first trimester placenta using dispase incubation of villous tissue at 4 °C, followed by a spontaneous cell release at 37 °C. After 24 h of culture, 90-95% of the cells were immunostained by anticytokeratin antibody, showing their epithelial characteristic. After 48 h of culture, these cells differentiated into syncytiotrophoblast, as shown by optic and electron microscopic study. The secretion of hCG, and of its free a and 13 subunits, and the secretion of hPL were studied as a function of cell culture time. While the level of secreted hCG and its free subunits was stable during 72 h of culture, the hPL level was undetectable during the first 48 h of culture, increasing continuously afterwards. Addition of dibutyryl cAMP from the start or after 96 h of cell culture induced an increase of hCG production and of its free subunits and also stimulated the secretion of hPL. This suggests that these cells maintained the capacity to respond to stimuli which increased intracellular cAMP level. Such a cell culture is of interest in further determining the mechanisms of early gestation involved in the differentiation and growth of placental cytotrophoblasts, and in the regulation of their endocrine functions.

Connexin expression and gap junctional intercellular communication in human first trimester trophoblast

“…Later, groups of cytotrophoblastic cells in close apposition were observed, consistent with an aggregation stage. Finally, after 72 h, large cellular masses with central nuclei mount and expanding cytoplasm were seen, thus confirming earlier studies (Dodeur et al, 1990;Tarrade et al, 2001). Only Cx43 was detected in trophoblastic cells cultured on plastic dishes.…”

Zika virus productively infects primary human placenta-specific macrophages

“…Therefore, our data appear to suggest that virus replication is inhibited after entry within this cell population in vitro. Primary human cytotrophoblasts do undergo spontaneous cell-to-cell fusion when left in culture for longer than 48 hours (14); therefore, analyses of later time points with these cultured cells may reflect a mixed population of cytotrophoblasts and syncytiotrophoblasts.…”

La placentation humaine

“…La compréhension des mécanismes impliqués dans la différenciation du cytotrophoblaste villeux en syncytiotrophoblaste a largement bénéficié de la possibilité d'isoler, de purifier et de maintenir ces cellules en culture [3][4][5]. La fusion des cytotrophoblastes est modulée in vitro par divers facteurs solubles, d'origine placentaire ou déciduale tels que l'hCG (hormone chorionique gonadotrope), des facteurs de croissance ( EGF : Epidermal Growth Factor, CSF : Colony-Stimulating Factor) [6] et des cytokines ( TGF-β : Transforming Growth Factor _, LIF : Leukocyte migration Inhibition Factor) [7,8], l'oestradiol [9], la pression partielle en oxygène [10] et l'expression d'une protéine rétrovirale, la syncytine [11,12], ou de jonctions communicantes dont la connexine 43 [13,14].…”