OBJECTIVETo develop a convenient method for isolation and purification of human extravillous cytotrophoblasts (EVCTs) and decidual stromal cells (DSCs) and establish a co-culture system.

METHODSThe DSCs were digested with trypsin and purified by Percoll gradient. The EVCTs were digested with trypsin and purified by BSA gradient. Immunohischemistry and immunofluorescent study are performed to characterize these isolated cells. The EVCTs and DSCs were placed in Matrigel-coated Transwell upper and lower chamber, respectively, to study the invasive ability of the EVCTs.

RESULTSImmunohischemistry revealed that the purity of EVCTs and DSC exceeded 95%. Cultured EVCTs retained their capacity to invade Matrigel-coated Transwell filters with the invasion index of 3.22-/+0.04.

CONCLUSIONThis co-culture model established by isolating highly purified EVCTs and DSCs in vitro can be useful for studying the trophoblast invasion mechanisms.

Cell Communication ; physiology ; Cells, Cultured ; Chorion ; cytology ; Coculture Techniques ; Decidua ; cytology ; Female ; Humans ; Models, Biological ; Stromal Cells ; cytology ; Trophoblasts ; cytology ; physiology

This study focused on the characterization of mesenchymal stromal cells (MSCs) from the chorion of human full term placenta from 15 donors. Chorionic MSCs revealed homologous fibroblast-like morphology and expressed CD73, CD29, CD105, and CD90. The hematopoietic stem cell markers including HLA DR, CD11b, CD34, CD79a, and CD45 were not expressed. The growth kinetics of their serial passage was steady at the later passages (passage 10). The multilineage capability of chorionic MSCs was demonstrated by successful adipogenic, osteogenic and chondrogenic differentiation and associated gene expression. Chorionic MSCs expressed genes associated with undifferentiated cells (NANOG, OCT4, REX1) and cardiogenic or neurogenic markers such as SOX2, FGF4, NES, MAP2, and NF. TERT was negative in all the samples. These findings suggest that chorionic MSCs undifferentiated stem cells and less likely to be transformed into cancer cells. A low HLA DR expression suggests that chorionic MSCs may serve as a great source of stem cells for transplantation because of their immune-privileged status and their immunosuppressive effect. Based on these unique properties, it is concluded that chorionic MSCs are pluripotent stem cells that are probably less differentiated than BM-MSCs, and they have considerable potential for use in cell-based therapies.
Antigens, CD/genetics/metabolism ; Cell Differentiation ; Cell Proliferation ; Cells, Cultured ; Chorion/cytology ; Female ; Gene Expression Regulation ; HLA-DR Antigens/genetics/metabolism ; Humans ; Mesenchymal Stromal Cells/*cytology/metabolism ; Placenta/*cytology ; Pregnancy ; Transcription Factors/genetics/metabolism

Endostatin, a carboxyl-terminal fragment of collagen XVIII is known as an anti-angiogenic agent, that specifically inhibits the proliferation of endothelial cell and the growth of several primary tumor. We report here the purification and characterization of the recombinant murine endostatin (rmEndostatin) which was expressed in a prokaryotic expression system. This rmEndostatin has similar physiochemical properties of yeast-produced recombinant endostatin, and it also specifically inhibits the proliferation and migration of bovine capillary endothelial cells stimulated by basic fibroblast growth factor. The biological activity of rmEndostatin was also shown by its anti-angiogenic ability on the chorioallantoic membrane of chick embryo in vivo. In this article, we demonstrate the refolding and purification of rmEndostatin, expressed using E. coli system, to a biologically active and soluble form. In addition, these results confirm the activity of endostatin as a potent anti-angiogenic agent. Copyright 2000 Academic Press.
Angiogenesis Inhibitors/pharmacology* ; Angiogenesis Inhibitors/isolation & purification ; Angiogenesis Inhibitors/genetics* ; Animal ; Blotting, Western ; Cattle ; Cell Movement/drug effects ; Chick Embryo ; Chorion/pathology ; Chorion/drug effects ; Circular Dichroism ; Collagen/pharmacology* ; Collagen/isolation & purification ; Collagen/genetics* ; Electrophoresis, Polyacrylamide Gel ; Endothelium, Vascular/drug effects ; Endothelium, Vascular/cytology ; Escherichia coli/genetics* ; Fibroblast Growth Factor, Basic/pharmacology ; Mice ; Neovascularization, Physiologic/drug effects ; Peptide Fragments/pharmacology* ; Peptide Fragments/isolation & purification ; Peptide Fragments/genetics* ; Protein Folding ; Recombinant Proteins/pharmacology ; Recombinant Proteins/isolation & purification ; Recombinant Proteins/genetics ; Solubility ; Yeasts/genetics