Comparison Of Biological Properties Between Human Peripheral Blood Derived Smooth Muscle Progenitor Cells And Mature Vessel Wall Smooth Muscle Cells

Objective:Compared the cell morphology and proliferation potential of smooth muscle progenitor cells(SPCs) derived from human peripheral blood with mature vessel wall smooth muscle cells(SMCs) to provide a reliable source of seed cells.Methods:1. Used Ficoll density gradient centrifugation to separate the mononuclear cells from adult peripheral blood. Cultured in the fibronectin-coated culture flask,and add EGM-2 medium which contails platelet-derived growth factor(PDGF-BB).Observed the morphological changes, liquid passage, cryopreservation and resuscitation by conventional methods.2. Obtained experimental vena saphena magna 3cm through the surgery, uses the collagen proteinase digestion, the difference to paste the wall law original generation of raise, raise in the EGM-2 culture medium including PDGF-BB. observation the change of cellular form, obtains the smooth muscle cell.3. Smooth muscle progenitor cells and smooth muscle cells were cell-climb filmed to detect the expression of a-SMA and calponinl by immunofluorescence.4. Reverse transcription-polymerase chain reaction detected the mRNA expression of smooth muscle actin (a-SMA) and calponinl.5. MTT detected the sixth generation proliferation of the smooth muscle progenitor cells and smooth muscle cell.Results:1. SPCs culture passage up to 80-90% confluence, appearing spiral arrangement,and presenting typical "peak, valley" form, sign smooth muscle cell morphology.2. Smooth muscle progenitor cells can express smooth muscle cell specific marker a-SMA and calponinl.3. The sixth generation of SPCs (n=10, A=0.43±0.02) proliferation was significantly faster than the same generations of SMCs (n=10, A=0.21±0.02, P <0 .01). Conclusion:1.Peripheral blood mononuclear cells in the stimulus of platelet-derived growth factor can differentiate into smooth muscle progenitor cells.2.Compared to smooth muscle cells, smooth muscle progenitor cells proliferate better, and not to see the aging in 3 months cultivation. Smooth muscle progenitor cells has the development of the possibility which one kind of ideal blood vessel organization project seed cell originates.