[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.

RESULTS AND CONCLUSION

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.