Differentiation of bone marrow mesenchymal stem cells into nucleus pulposus-like cells after co-culture with nucleus pulposus cells

Wei Ge-jin; Huang Yu-qiang; Qin Wan-an; Liao Chang-chuan; Lin Zhou-dan

doi:10.3969/j.issn.2095-4344.2013.45.003

Abstract：BACKGROUND:Bone marrow mesenchymal stem cells in recent years have been widely used as seed cells for nucleus pulposus tissue engineering. Appropriate extracellular microenvironment and growth factors are required for the induction of bone marrow mesenchymal stem cells differentiating into nucleus pulposus cells. <br> OBJECTIVE:To observe the differentiation of rabbit bone marrow mesenchymal stem cells into nucleus pulposus-like cells in vitro induced by biological inducers and co-culture conditions. <br> METHODS:Rabbit bone marrow mesenchymal stem cells and nucleus pulposus cells were isolated and cultured using monolayer culture method, respectively. Passage 3 bone marrow mesenchymal stem cells were placed in the Transwel culture plates for co-culture with the passage 3 nucleus pulposus cells and then were induced biological y to differentiate into the nucleus pulposus-like cells for 21 days. Bone marrow mesenchymal stem cells were cultured alone in the high-glucose Dulbecco’s modified Eagle’s medium as negative controls. <br> RESULTS AND CONCLUSION:The expressions of CD29 and CD44 were positive and CD34 and CD45 were negative in the bone marrow mesenchymal stem cells. After 21 days of co-culture with nucleus pulposus cells, the morphology of bone marrow mesenchymal stem cells changed obviously from polygonal to oval shape, and col agen type Ⅱ granules were increased significantly. Reverse transcription-PCR results showed that expression of col agen type Ⅱ mRNA was significantly increased after induction. These findings indicate that bone marrow mesenchymal stem cells can be induced into differentiate into nucleus pulposus-like cells in co-culture plus biological inducer conditions in vitro.