Experimental studies on induction of rat bone marrow endothelial progenitor cells

Objective:To culture and induce endothelial cells from rat bone marrow in vitro and preparation for the vascular tissue engineering research. Method: Bone marrow cell suspension of adult rat was prepared by discontinuous gradient centrifugation on Ficoll(density 1.077 g/ml), mononeuclear cells in the middle layer were collected and put into selective culture medium and cultured in the fibernectin coated plate. The cells were observed under inverted microscope every day. The cells were identified by transmission electron microscope , flow cytometer and immunohistochemical staining with factor Ⅷ, CD31, Lectin antigen. Result: The cells had round or spindle shape. The cells were factor Ⅷ, Lectin and VEGFR positive. Under transmission electron microscope, the cells displayed Weibel-Palade bodies which was the character of endothelial cells. Conclusion: Rat bone marrow EPCs are cultured and induced successfully in vitro in this study. Discontinuous density gradient centrifugation is effective approach to isolate rat bone marrow endothelial progenitor cells. The cells induced in vitro have the same characters as endothelial cells.