[Isolation, culture and chondrogenic differentiation of goat bone marrow mesenchymal stem cells].

OBJECTIVE

To explore the isolation, in vitro culture and chondrogenic differentiation of goat bone marrow mesenchymal stem cells (BMSCs).

METHODS

Bone marrow was harvested from a 10-month-old Chinese goat for adherent culture of the BMSCs in vitro. Flow cytometry was performed to detect the cell surface markers of the BMSCs of the fourth generation. The induction medium (containing 10% fetal bovine serum, high-glucose DMEM, 6.25 microg/ml insulin, 6.25 microg/ml transferrin, 50 microg/ml vitamin C, 100 nmol/L DXM and 10 ng/ml transforming growth factor-beta1) was then applied for chondrogenic differentiation. Cytochemical staining, RT-PCR and Western blotting were performed to detect the expressions of type II collagen and aggrecan in the cells at the time points of 0, 1, 2 and 4 weeks.

RESULTS

The goat BMSCs grow well in vitro with a high purity in the fourth generation. The expression of chondrocyte phenotypes were observed at 1, 2 and 4 weeks, which became more obvious as the culture prolonged. The mRNA and protein expression of type II collagen and aggrecan in the BMSCs increased obvious after the induction and had reached a satisfactory level by 2 weeks.

CONCLUSION

Goat BMSCs have the potential to differentiate into chondrocytes in vitro, and the results of this study provide the experimental basis for application of goat BMSCs in bone and cartilage tissue engineering in vivo.