Establishment and Biological Characterization of an Asian Human First Trimester Placental Villus-derived Trophoblastic Cell Line (HPT-8) in Vitro

Objective: To establish the human first trimester placental villus-derived trophoblastic cell line(HPT-8) in vitro and to analysis its biological characterization of the cell line in Asian.Methods:The placental villus were digested by pancreatic enzyme and collagenase I.Primary culture of human trophoblast cells was carried out by combined digestion,and the subsequent serial passage of the cells performed by trypsin digestion.The HPT-8 in vitro was identified and assayed by light microscope,scanning electron microscope(SEM),transmission electron microscope(TEM),immunohistochemistry(IH) and immunofluorescence laser confocal microscopy.Results:During 5～7 passage numbers of the cell primary culture in vitro,most cells have apoptosis phenomenon.A few trophoblastic cells became the immortalization cell.Endocytoplasmic reticulum distended expressly.There was desmosome structure between the trophoblast cells.Keratin 18 and vimentin specifically bound to the cytoplasm of trophoblastic cells with direct immunofluorescence double marker.Keratin 7,motility related protein-1,epidermal growth factor receptor,stromal cell derived factor and placental alkaline phosphatase were specifically bound to the cytoplasm or cytolemma of the cells with IH.But human chorionic gonadotropin and placental lactogen didn't find in the cytoplasm of HPT-8 with IH.Conclusions: We gained pure trophoblastic cell used combined digestion with pancreatic enzyme and collagenase I.The study of light microscope,SEM,TEM,IH and immunofluorescence laser confocal microscopy detection confirmed that the HPT-8 in vitro was established from appearance,cystoskeleton and functional parameter.The cell line model with the part function of endocrine secretion could evolve empirical study in the future.