A protocol for the preparation of highly purified human placental cytotrophoblast cells from normal first trimester placental villi is described. Cytotrophoblast cells were isolated from placentas (6 - 14 weeks of gestation) by a process of mincing, sequential two-step enzymatic treatment of placental villi by collagenase, hyaluronidase, DNAse, and trypsin, followed by discontinuous Percoll gradient centrifugation, and immunomagnetic separation employing antibodies to CD45 to remove contaminating leukocytes. A population of 96 % pure living mononuclear cytotrophoblast cells (identified by positive cytokeratin 8,18 and E-cadherin staining) was obtained at a density of 1.048 and 1.062 g/ml Percoll. The isolated cells were characterized employing routine histology, conventional immunocytochemistry, fluorescence confocal laser scanning microscopy and flow cytofluorometry for the cell antigens CK 8,18, b-hCG, hPL, E-cadherin, integrin subunits a1, a5, a6, av, b1, b3, b4, and possess the morphological and structural characteristics of cytotrophoblast cells. The presented method of cytotrophoblast isolation should facilitate ongoing in vitro studies of trophoblast adhesion, differentiation, migration, proteolytic activity, and invasion, and clarify how these events differ from those of malignant cells.     «

A protocol for the preparation of highly purified human placental cytotrophoblast cells from normal first trimester placental villi is described. Cytotrophoblast cells were isolated from placentas (6 - 14 weeks of gestation) by a process of mincing, sequential two-step enzymatic treatment of placental villi by collagenase, hyaluronidase, DNAse, and trypsin, followed by discontinuous Percoll gradient centrifugation, and immunomagnetic separation employing antibodies to CD45 to remove contaminatin...     »

Ein Isolierungsprotokoll für humane Zytotrophoblastzellen aus normalen Plazenten des ersten Trimenons wird beschrieben. Die Zytotrophoblastzellen werdem mittels mechanischer Zerkleinerung, sequentieller enzymatischer Auflösung, Percollgradientenzentrifugation und immunomagnetischer Separation isoliert. Man erhält eine 96%-ig reine, vitale Zytotrophoblastzellpopulation. Die isolierten Zellen werden anhand von Routinehistologie, konventioneller Immunozytochemie, konfokaler Laserscan Fluoreszenzmikroskopie und Durchflußzytometrie für folgende Zellantigene charakterisiert: CK 8/18, β-hCG, hPL, E-Cadherin, Integrinuntereinheiten α1, α5, α6, αv, β1, β3, β4. Die isolierten Zellen können morphologisch und strukturell als Zytotrophoblastzellen identifiziert werden.     «

Ein Isolierungsprotokoll für humane Zytotrophoblastzellen aus normalen Plazenten des ersten Trimenons wird beschrieben. Die Zytotrophoblastzellen werdem mittels mechanischer Zerkleinerung, sequentieller enzymatischer Auflösung, Percollgradientenzentrifugation und immunomagnetischer Separation isoliert. Man erhält eine 96%-ig reine, vitale Zytotrophoblastzellpopulation. Die isolierten Zellen werden anhand von Routinehistologie, konventioneller Immunozytochemie, konfokaler Laserscan Fluoreszenzmik...     »