| Purpose Our research is to study the ability of Chondrogenic Differentiation of Rat Adipose-derived Stem Cells by isolating Chondrogenic Differentiation of Rat Adipose-derived Stem Cells by isolating and culturing in vitro and to observe the influences of different concentrations of TGF-β1 on proliferation and differentiation, in order to get the foundation forMethods The adipose tissue were isolated from the inguinal fat pads of the three-weeks old Sprague-Dawley rat, then digesed with collagenase-I and placed in Dulbeco Modified Eagle Medium(DMEM)with fetal bovine serum in primary cultivation.We observed the growth and appearance of the cells by a inversion microscope. Take the third generation of ADSCs into 5 groups, including a control group(ADSCs medium:high glucose DMEM,10% fetal bovine serum,50 nmol/L ascorbic acid,6.25 mg/L insulin, 100U/ml penicillin, 100μg/ml streptomycin 100nmol/L dexamethasone, without TGF-β-1) and 4 groups of TGF-β1 concentrations(respectively 1,10,40,80μg/L increase of ADSCs medium). Both of the proliferation and differentiation of ADSCs was examinated by MTT test,and chondrogenic differentiation of ADSCs was examinated by II collagen immunohistochemical staining and toluidine blue staining.Results The primary cells began to adherence at the 4-6th hour after settlement.They proliferative quikely at the 7-8th day,arranging according to a certain direction,such as whirlpool or bundle.The cells proliferatived quick and steady after passaging. These cells stained with toluidine blue and type II collagen immunohistochemical staining showed:10μg/L group positive; 40μg/L group and 80μg/L group, weakly positive; 0μg/L group and 1μg/L group was negative. MTT test showed:10μg/L TGF-β1 can promote cell proliferation significantly (P <0.05). Conclusions Rat Adipose-derived Stem Cells can be induced to differentiate into cartilage cells; 10μg/L is the optimal concentration of TGF-β1 for Adipose-derived Stem Cells proliferating and differentiating to cartilage,and 0,1,40,80μg/L were not suitable.
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