In vitro differentiaion of peripheral blood mononuclear cells into smooth muscle progenitor cells / 上海第二医科大学学报

ABSTRACT

Objective To optimize methods of culturing smooth muscle progenitor cells(SPCs) from mononuclear cells(MNCs) of peripheral blood. Methods Human MNCs isolated from buffy coat were seeded on M-199 with bovine pituitary extraction.On the eighth day outgrowth cells were stimulated with platelet-derived growth factor-BB(PDGF-BB).Fifteen days later,immunofluorescence,Western blot or RT-PCR was used to analyzed the expression of smooth muscle cell specific ?-actin(?-SMA),smooth muscle myosin heavy chain(SM MHC),Calponin,CD34,Tie-2 and Flk-1,and fluorescence activated cell sorter was employed to examine ?-SMA positive cells ratio. Results The cells stimulated by PDGF-BB for 15 d were positive for ?-SMA,SM MHC,Calponin,CD34 and Flk-1,but negative for Tie-2.The ?-SMA positive cells ratio was(90.57?5.63)%,significantly different from that of the control(P