Primary culture of bovine retinal capillary endothelial cells and pericytes in vitro
- VernacularTitle:牛视网膜微血管内皮细胞和周细胞的体外培养
- Author:
Xin XIA
;
Xun XU
;
Qing GU
- Publication Type:Journal Article
- Keywords:
Cell culture;
Pericyte;
Endothelium, vascular;
Retinal vessels
- From:
Chinese Journal of Ocular Fundus Diseases
1996;0(01):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective To probe a selective cultural method for bovine retinal endothelial cells (BREC) and pericytes (BRP) in vitro. Methods With the isolation of active retinal blood vessels, BREC were cultured in a fibronectin coated substrate and Dulbecco′s Modified Eagle′s Medium (DMEM) supplemented with 10% human serum and 100 ?g/ml heparin, while homogeneous cultures of retinal pericytes were obtained when isolated microvessels were seeded to uncoated dishes and grown in DMEM supplemented with 20% fetal bovine serum. BREC were identified by acetylated-low density lipoprotein (Dil-Ac-LDL) incorporation and immunohistochemical method of Von Willebrand factor, while BRP were identified by immunohistochemical method of ?-isoform of smooth-muscle actin. Results The purity of selectively cultured BREC and BRP was more than 98%, being reproducible. BREC got together around the microvessel fragments with the small-cyprinoid-like configuration at first,and could phagocytize Dil-Ac-LDL with the expression of fluorescence in cytoplasm. The expressions of Von Wllebrand factor and ?-isoform of smooth-muscle actin were positive and negative respectively in BREC, while were negative and positive respectively in BRP. Conclusion BREC and BRP with high purity can be obtained by using selective culture and coated-dishes respectively which are simple and repeatable methods.
