[In vitro study on multiple differentiation potential of swine synovium-derived MSCs].
OBJECTIVE
To study the method of isolating and culturing synovium-derived MSCs (SMSCs), and to investigate its multiple differentiation potential in vitro.


METHODS
Three 2-month-old Changfeng hybrid swines weighing 8-10 kg (male and female) were used. SMSCs were harvested from the synovium of swine knee joints and cultured in vitro. When the SMSCs at passage 3 reached confluence, basic culture medium was removed, and the multiple differentiation potential of SMSCs was demonstrated in specific induction media (experimental group). The cells at passage 3 cultured with basic culture medium served as control group. After 21 days of chondrogenic differentiation, the cells underwent toluidine blue staining, immunohistochemistry staining and real-time fluorescence quantitative PCR detection. After 10 and 21 days of osteogenic differentiation, the cells underwent ALP staining and Alizarin red staining, respectively. After 21 days of adipogenic differentiation, the cells underwent Oil red O staining.


RESULTS
SMSCs displayed long and thin or polygonal morphology 24 hours after culture. They proliferated fast 48 hours after culture and presented large number of spindle-shaped cells with few globular cells 72 hours after culture. For the experimental group 21 days after chondrogenic induction, the cells were positive for toluidine blue staining with the formation of Aggrecan outside the cells; the immunohistochemistry staining revealed the expression of Col II; the real-time fluorescence quantitative PCR detection showed that the expressions of Col II A1, Aggrecan and SOX9 mRNA of the experimental group were greater than that of control group (P < 0.05). The cells were positive for ALP staining 10 days after osteogenic induction, and positive for Alizarin red staining 21 days after osteogenic induction, with the formation of calcium nodules. Oil red O staining displayed the formation of lipid droplets inside the cells 21 days after adipogenic induction. For the control group, the results of all the staining assays were negative except the ALP staining presenting with slight positive result.


CONCLUSIONS
SMSCs can be isolated from knee joint of swine and proliferate and differentiate into osteogenic, adipogenic and chondrogenic cells in vitro. SMSCs may be a promising source of seed cells for tissue engineering.
Wang
Yang
Y
Department of Orthopaedics, Shanghai Ninth People's Hospital, Medical College of Shanghai Jiaotong University, Shanghai, 200011, PR China.
Wang
You
Y
Rui
Yunfeng
Y
Du
Lin
L
Tang
Tingting
T
Dai
Kerong
K
D000818:Animals; D001854:Bone Marrow Cells / Q000166:cytology*; D002454:Cell Differentiation / Q000502:physiology*; D002478:Cells, Cultured; D005260:Female; D008297:Male; D059630:Mesenchymal Stem Cells / Q000166:cytology*; D010006:Osteoblasts / Q000166:cytology*; D013552:Swine; D013583:Synovial Membrane / Q000166:cytology*; D023822:Tissue Engineering / Q000379:methods