YANG Ping 1 , YIN Shuo 2 , CUI Lei 1 , LI Hong 2 , WU Yingchen 1 , LIU Wei 1 , CAO Yilin. 1
  • 1Department of Plastic and Reconstructive Surgery, the Ninth People’s Hospital, Affiliated to Shanghai Jiaotong University, Shanghai, 200011, P.R.China;;
  • 2Shanghai Tissue Engineering Research and Development Center.;
Tissue engineered blood vessel; Adipose derived stem cells; Smooth muscle cells; Cell induce
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To study the feasibil ity of human adipose derived stem cells (ADSCs) in monolayer culture induced into smooth muscle cells in vitro as seeding cells in vascular tissue engineering. Methods The mononuclear cells in human adipose were separated by collagenase treatment and seeded on culture dishes with the density of 5 × 105/cm2. Cells
were cultured in M-199 plus 10% FBS. When reaching confluence, the cells were subcultured by 0.1% trypsin and 0.02%
EDTA treatment, PDGF-BB (50 ng/mL) and TGF-β1 (5 ng/mL) were added at the passage 1 to enhance the smooth muscle cells’ phenotype. Cells were cultured under the inducing medium for 14 days. The morphology of induced cells was observed under the microscope. Cellular immunofluorescence and RT-PCR were used to determine the expression of smooth muscle cell markers of the post-induced cells. Flow cytometry (FACs) was used to examine the positive rate of induced team. Results Cocultured in M-199 media including TGF-β1 and PDGF-BB, the prol iferating capabil ity of the induced cells was significantly downregulated compared with the uninduced cells(P  lt; 0.01). The induced cells exhibited “Hill and Valley” morphology, while the uninduced cells were similar to ADSCs of P0 which had the fibroblast-l ike morphology. The results of immunofluorescence indicated that the induced cells expressed smooth muscle (SM) cell- specific markers including α-smooth muscle actin (α-SMA), SM-myosin heavy chain (SM-MHC) and Calponin. The results of RT-PCR revealed that the induced cells also expressed α-SMA, SM-MHC, Calponin and SM-22α.The positive rates of α-SMA, SM-MHC and Calponin in FACs were 3.26% ± 1.31%, 3.55% ± 1.6% and 4.02% ± 1.81%, respectively, before the cells were induced. However, 14 days after the cell induction, the positive rates were 48.13% ± 8.31%, 45.33% ± 10.68% and 39.13% ± 9.42%, respectively. The positive rates in induced cells were remarkably higher than those in uninduced cells(P  lt; 0.01). Conclusion The human ADSCs can be induced to express vascular smooth muscle markers, and they are a new potential source of vascular tissue engineering.

Citation： YANG Ping,YIN Shuo,CUI Lei,LI Hong,WU Yingchen,LIU Wei,CAO Yilin.. EXPERIMENT OF ADIPOSE DERIVED STEM CELLS INDUCED INTO SMOOTH MUSCLE CELLS. Chinese Journal of Reparative and Reconstructive Surgery, 2008, 22(4): 481-486. doi: Copy