Differentiation of SOX-9 and GDF-5 co-transfected bone marrow mesenchymal stem cells into nucleus pulposus cells

doi:10.3969/j.issn.2095-4344.2015.19.001

Abstract

Abstract:

BACKGROUND: Transplantation of mesenchymal stem cells to prevent and treat degeneration of the intervertebral disc is a feasible method. Mesenchymal stem cells co-transfected by SRY-related high mobility group-box gene 9 (SOX-9) and growth differentiation factor-5 (GDF-5) can differentiate into nucleus pulposus cells, in order to obtain greater effect of induction and proliferation of nucleus pulposus cells.
OBJECTIVE: To investigate the effect of SOX-9 and GDF-5 co-transfection on the differentiation of rabbit bone marrow mesenchymal stem cells into nucleus pulposus cells.
METHODS: We separated and cultured bone marrow mesenchymal stem cells from the bone marrow of rabbit aged 4 months. Passage 3 cells were divided into five groups and in vitro induced to differentiate into nucleus pulposus cells: non-transfected group, empty vector transfection group, SOX-9 transfection group, GDF-5 transfection group, SOX-9 and GDF-5 co-transfection group. At 14 days after transfection, RT-PCR was employed to assay SOX-9, GDF-5 and collagen type II mRNA expressions in bone marrow mesenchymal stem cells. The marker of nucleus pulposus cells-KRT19 expression was also detected by immunohistochemical staining.

RESULTS AND CONCLUSION: In the co-transfection group, the mRNA expressions of SOX-9, GDF-5, and collagen type II were significantly higher than those in the SOX-9 transfection group, GDF-5 transfection group, and both these two groups, respectively (P < 0.05). Cells were positive for KRT19 in the SOX-9 and GDF-5 groups, and strongly positive for KRT19 in the co-transfection group. These findings indicate that double gene-transfected bone marrow mesenchymal stem cells are better than single gene-transfected cells with regard to differentiation into nucleus pulposus cells and secretion of extracellular matrix.

Key words: Bone Marrow, Mesenchymal Stem Cells, Transfection, SOX9 Transcription

CLC Number:

R394.2

Du Zhi-cai, Yin He-ping, Li Shu-wen, Wu Hai-jun, Bai Ming, Cao Zhen-hua, Meng Ge-dong. Differentiation of SOX-9 and GDF-5 co-transfected bone marrow mesenchymal stem cells into nucleus pulposus cells[J]. Chinese Journal of Tissue Engineering Research, 2015, 19(19): 2953-2958.

Figures/Tables 3

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