[Isolation, identification and in vitro culture of human glomerular mesangial cells].
OBJECTIVE
To define the optimal condition to culture human glomerular mesangial cells in vitro.


METHODS
With a three-layer micropore filter device, we separated the glomerular mesangial cells from normal human, with were subjected to identification with immunohistochemical method. Fluorescein isothiocyanate (FITC) staining was performed for labeling of the monoclonal antibodies so as to identify the cells' superficial markers including factor VIII, keratin, cytokeratin, desmine, et al. Glomerular endothelial cells isolated from the same human tissues were also cultured in vitro under the same condition, serving as negative control.


CONCLUSIONS
The antibodies related to anti-factor VIII, and those of antikeratin and anticytokeratin were negative in the isolated glomerular mesangial cells, while the antibodies for desmine, anti-collagen IV, anti-fibronectin, and anti-laminin were positive, offering evidence for confirming the identification of the isolated and cultured mesangial cells.
Sun
Xiao-yan
XY
Department of Embryology and Histology, First Military Medical University, Guangzhou 510515, China. yanzisun1979@sina.com
An
Jing
J
Fu
Bo
B
Li
Jin
J
D002469:Cell Separation; D002478:Cells, Cultured; D024141:Collagen Type IV / Q000032:analysis; D003893:Desmin / Q000032:analysis; D005353:Fibronectins / Q000032:analysis; D005920:Glomerular Mesangium / Q000737:chemistry / Q000166:cytology*; D006801:Humans; D007150:Immunohistochemistry; D007797:Laminin / Q000032:analysis
D024141:Collagen Type IV; D003893:Desmin; D005353:Fibronectins; D007797:Laminin