Differentiation of Mouse BM-MNCs into Endothelial Cells in vitro

Objective:To culture and induce endothelial cells from mouse bone marrow in vitro and prepare for the vascular tissue engineering research.Methods: Bone marrow cell suspension of adult mouse was prepared by discontinuous gradient centrifugation on Ficoll(density 1.074g/ml),mononeuclear cells in the middle layer was collected,put into selective culture medium and cultured in the fibernectin coated plate.The cells were observed under inverted microscope every day.The cells was identified by transmission electron microscope,flow cytometer and immunohistochemical staining with factor Ⅷ、CD31、Lectin antigen.Results: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 were endothelial character.Conclusion:This study cultures and induces mouse bone marrow EPCs successfully in vitro.Discontinuous density gradient centrifugation is effective approach to isolating mouse bone marrow stromal cells.The cells induced in vitro have the same characters as endothelial cells.