Isolation and culture of term human trophoblast cells.
Experimentation with most human cell types is restricted to the use of cell lines, and this limits our ability to extrapolate interpretations to the in vivo condition. However, in studying human trophoblast cells, we have a unique opportunity to obtain large quantities of readily available human tissue. In this chapter, we outline the methodology for purification of human trophoblast cells from term placentas. The procedures are based on enzymatic dissociation of villous placental tissue, followed by gradient centrifugation and immunomagnetic bead purification. Purity may be assessed by immunocytochemistry or flow cytometry using a number of markers to identify both cytotrophoblast cells and cellular contaminants. The resulting cytotrophoblast cell populations have excellent viability and purity, and may be subjected to long-term culture.
Petroff
Margaret G
MG
Department of Anatomy and Cell Biology, University of Kansas Medical Center, Kansas City, USA.
Phillips
Teresa A
TA
Ka
Hakhyun
H
Pace
Judith L
JL
Hunt
Joan S
JS
D018929:Cell Culture Techniques / Q000379:methods*; D002469:Cell Separation / Q000379:methods*; D002470:Cell Survival; D002478:Cells, Cultured; D002824:Chorionic Villi; D005260:Female; D006801:Humans; D018189:Immunomagnetic Separation; D010920:Placenta / Q000166:cytology; D011247:Pregnancy; D047929:Term Birth; D014327:Trophoblasts / Q000166:cytology*