Abstract:

Objective To improve the method for primary cultivation of rat pulmonary microvascular endothelial cells (PMVECs), and identify the cultivated cells. Methods Method for primary cultivation of rat PMVECs was improved from the procedures of animal selection, sampling, perfusion and tissue piece pasting. After subculturing, freezing and thawing, PMVECs were observed by inverted microscopy for morphology and growth behavior, identified by immunohistochemical staining for expression of factor Ⅷ and CD31, and observed by fluorescence microscopy for binding with lectin from BSI (FITC-BSI binding assay). Results Under inverted microscope, the cultivated cells exhibited as polygon or fusiform, and displayed typical cobblestone-like morphology after fusion to monolayer, with contact inhibition. Immunohistochemical staining revealed that the expression of factor Ⅷ and CD31 was positive. Besides, there were positive findings for FITC-BSI assay. Conclusion The improved method for primary cultivation is easy to handle, with favourable repeatability and success rate, and the morphological and biological characteristics of cultivated cells are in line with PMVECs.

Key words: pulmonary microvascular endothelial cells, cell cultivation, cell identification