Growth differentiation factor 5 induces adipose-derived stem cells differentiating into chondrocytes / 中国组织工程研究

ABSTRACT

BACKGROUND:Growth differentiation factor 5 (GDF5) has been shown to play a crucial role in the development of chondrocytes via different regulatory mechanisms. OBJECTIVE:To explore the effect of GDF5 on the chondrogenic differentiation of adipose-derived stem cel s (ADSCs). METHODS:ADSCs in passage 4 isolated from Japanese White rabbits were cultured in the medium containing 0 (blank control group), 10, 50, 100, 150 and 200μg/L, respectively. The morphological changes were observed, and the optimal concentration of GDF-5 was screened, and its round coverslips at 1, 2 and 3 weeks of culture were selected to undergo immunocytochemistry, toluidine blue staining and immunofluorescent staining. RESULTS AND CONCLUSION:The growth of ADSCs was stable in the medium containing 10 and 50μg/L GDF5, while apoptotic cel s appeared after induction with 200μg/L GDF5. In the medium containing 100 and 150μg/L GDF5, some spindle-shaped cel s changed into irregular shape, and became round or oval after 7-10 days of culture. The optimal concentration of GDF5 was 100μg/L. The cel s transfected by 100μg/L GDF5 were positive for type II col agen obviously and with blue-stained nucleus. The transfected cel s were positive for toluidine blue, and metachromatic granules were visible in the cytoplasm. The proteoglycan mRNA expression of the transfected cel s was significantly increased, and reached the highest at 3 weeks. These results suggest that GDF5 promotes the chondrocytic differentiation of ADSCs.