Abstract: To investigate and establish the methods of in vitro culturing and identification of endothelial progenitor cells (EPCs) of Bama Minipigs. The mononuclear cells were gained from Bama Minipigs bone marrow by density gradient centrifugation，inoculated （2×10⁷） in the culture plates，which were coated with HFN and induced differentiation to EPCs.The EPCs were observed by inverted microscope, compare the proliferation status of both derived EPCs by observing the growth curve，MTT method，and the express of EPCs-related antigen ac-LDL, UEA-1, AC133 and vWF by immunofluorescence staining.MTT results showed that the proliferation of the cells were obvious at 3 to 5 d.FACs test results showed that vWF adherent cells twice percentage of positive cells was 74.91%, AC133-positive cell percentage was 79.54%, while the secondary without adherent cells induced vWF and AC133-positive rate were 33.65% and 39.87%. The cells were identified by immunofluorescence expression of ac-LDL, UEA-1, AC133 and vWF were positive. Thus, the cells isolated and cultured from Bama Minipigs bone marrow were shown that strong ability of self-renewal proliferation and multipotency of differentiation which identified as the endothelial progenitor cells.

Key words: Bama Minipigs; bone marrow; endothelial progenitor cells; isolation and culture