Le Bellego, Frédérique; Vaillancourt, Cathy 
ORCID: https://orcid.org/0000-0003-0543-6244 et Lafond, Julie
(2009).
Isolation and culture of term human cytotrophoblast cells and in vitro methods for studying human cytotrophoblast cells' calcium uptake
In:
Human Embryogenesis: Methods and Protocols.
Methods in Molecular Biology, 550
(550).
Humana Press, Totowa, NJ, pp. 73-87.
Résumé
Human primary cytotrophoblast cell culture is a very useful model to study the endocrine and immunological functions of syncytiotrophoblasts, as well as the ion exchange between the mother and her fetus, like calcium. In this chapter, we expose the procedure to (1) isolate and purify the cytotrophoblast cells from human term placenta and (2) study syncytiotrophoblast calcium uptake. First, the methodology is based on the enzymatic dissociation of villous placental tissue, followed by Percoll gradient separation. Purity is assessed by flow cytometry using staining against cytokeratin-7, protein specific for trophoblast cells. Cell proliferation is evaluated by a Thiazolyl Blue Tetrazolium Bromide (MTT) assay, hormonal secretion is measured by enzyme-linked immunosorbent assay (ELISA), and fusion is estimated by immunofluorescence using staining against desmosomal proteins. Second, we describe the calcium uptake experiment using the cytotrophoblast cells in culture.
| Type de document: | Chapitre de livre |
|---|---|
| Mots-clés libres: | - |
| Centre: | Centre INRS-Institut Armand Frappier |
| Date de dépôt: | 30 juin 2024 21:51 |
| Dernière modification: | 30 juin 2024 21:51 |
| URI: | https://espace.inrs.ca/id/eprint/14874 |
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