[Connective tissue growth factor induced differentiation of placenta mesenchymal stem cell into dermal fibroblast].
OBJECTIVE
To investigate the possibility of placenta mesenchymal stem cells (PMSCs) differentiation into dermal fibroblast, and the potency of PMSCs used in cutaneous wound healing and stored as seed cells.


METHODS
Enzyme digestion method was used to obtain PMSCs, and PMSCs were amplified after culture in vitro. Flow cytometry assay, osteogenic and adipogenic differentiation were done for MSCs identification. The induction medium composed of DMEM/F12 + 50 microg/ml VC + 100 ng/ml connective tissue growth factor (CTGF) was added into the 24-well plate for 16 days induction period. Pictures were taken to record morphologic change. Immunofluorescence tests were performed to detect Vimentin, FSP-1, collagen I , collagen III, desmin and laminin expression before and after induction. At the same time osteogenic and adipogenic differentiation were used to assay the differentiation ability change after induction. The induced dermal fibroblasts were frozen in liquid nitrogen and recovery and trypan blue was used to detect cell viability.


RESULTS
After CTGF induction, PMSCs got obvious fibroblasts morphology, the protein level of Vimentin, FSP-1, collagen I, collagen III and Laminin increased, PMSCs started to express Desmin, the dermal fibroblasts specific proteins, and osteogenic and adipogenic differentiation ability was diminished. PMSCs were successfully induced into dermal fibroblasts, and these induced cells could get a high cell viability ( more than 90% ) after recovery.


CONCLUSIONS
PMSCs could be induced into dermal fibroblasts by CTGF in vitro. PMSCs have the potential application in skin wound healing, and can be used as seed cells of dermal fibroblasts.
Department of Burns and Orthopedics, the First People's Hospital of Huai'an, Huai'an 223200, China.
Wei-Xi
WX
Yang
Xi
X
Ouyang
Yun-Qing
YQ
Song
Xiao-Guang
XG
Zhang
Jun
J
Zhang
D002454:Cell Differentiation / Q000187:drug effects*; D002478:Cells, Cultured; D055513:Connective Tissue Growth Factor / Q000494:pharmacology*; D005260:Female; D005347:Fibroblasts / Q000166:cytology* / Q000187:drug effects; D006801:Humans; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000187:drug effects; D010920:Placenta / Q000166:cytology; D011247:Pregnancy
D055513:Connective Tissue Growth Factor