In Vitro Experiments Of Adipose Stem Cells Induced Into Vascular Smooth Muscle Cells

Objective:To investigate the feasibility of adipose derived stem cells in intro differentiate into vascular smooth muscle cells with transforming growth factor beta 1 and platelet derived growth factor BB,find the new source of seeding cells to construct the tissue-engineered vessels。Method:The original adipose stem cells in human adipose were separated by collagenase treatment,To separate the cells of CD34+ by MACS,adherent cells were allowed to grow to near confluency。After grow 24 hours,the CD34+were separated two groups:In experimental group,PDGF-BB(50ng/ml)and TGFβ1 (2.5ng/ml)was added to the 5%M199 medium to induce ASCs to vSMCs, Control group were cultured by the 5%M199 medium,After 14 days,cell morphology was observed under inverted phase contrast microscope。Methods of immunoflurescence staining,and RT-PCR were used to determine the expression of smooth muscle cell specific markers(α-SMA,Calponin,SM22α,SM-MHC) of the experimental cells。FACS was used to examine the positive rate of experimental group cells' markers。Results:When ASCs were cultured in 5%M199 medium containing PDGF-BB (50ng/ml)and TGFβ1(2.5ng/ml),cells showed "peak and valley" growth pattern specified as mature VSMCs and began to express VSMC's markers asα-SMA,Calponin,SM 22α,SM-MHC;FACS showed that the positive percentage ofα-SMA,Calponin,SM-MHC of experimental cells were 5.11±0.44%,5.73±0.54%,8.42±0.47%。Alter 14 days,the positive percentage 50.58±2.71 %,42.94±2.76%,49.31±0.67%and the positive percentage had siginificently increasing,have a significant difference between them。Conclusion:ASCs were induced in vitro by specific environment after a period of time,the cells show the significant characteristics of VSMCs and has the potentiality to construct the tissue-engineered vessels as seeding cells。...