LIM mineralization protein expression in LIM mineralization protein-1 gene-transfected bone marrow mesenchymal stem cells

doi:10.3969/j.issn.2095-4344.2014.23.005

Abstract

Abstract:

BACKGROUND: In vitro experiments suggest that LIM mineralization protein-1 (LMP-1) gene can increase the expression of LMP-1 protein in osteoporotic bone marrow mesenchymal stem cells.

OBJECTIVE: To investigate the LMP-1 expression in osteoporotic bone marrow mesenchymal stem cells transfected with RV-LMP-1-GFP in vitro.

METHODS: Twelve SD female rats were selected and subjected to bilateral ovariectomy for establishment of osteoporosis models. After 2 months of feeding, bilateral femurs, tibiae, and humeri of rats were taken to isolate and culture bone marrow mesenchymal stem cells. Passage 3 cells were taken and randomly divided into ovariectomized group and LMP-1 transfection group. Another six rats only underwent removal of the same amount of fat tissue around the ovary, and passage 3 cells which were harvested as those in the former two groups served as sham group. RT-PCR and western blot assay were performed to determine the expression of LMP-1 protein and mRNA.

RESULTS AND CONCLUSION: Under an inverted fluorescence microscope, transfected bone marrow mesenchymal stem cells from osteoporotic rats showed green fluorescent expression. The three groups all could express LMP-1 at the protein and mRNA levels. The expression levels of LMP-1 protein and mRNA in the LMP-1 transfection group were significantly higher than those in the ovariectomized group and sham group (P < 0.05), but there was no statistical difference between the latter two groups (P > 0.05). The successful expression of the RV-LMP-1-GFP gene in osteoporotic SD rat bone marrow mesenchymal stem cells was realized at the protein and mRNA levels; moreover, the expression of LMP-1 was increased dramatically. These findings indicate that LMP-1 gene is successfully transferred into osteoporotic SD rat bone marrow mesenchymal stem cells, and significantly elevates the expression of LMP-1 mRNA and protein.

中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程

全文链接：

Key words: stem cells, mesenchymal stem cells, retroviridae, osteoporosis

CLC Number:

R394.2

Jin Xia-sheng, Wang Zi-jiang, Xiang Chuan. LIM mineralization protein expression in LIM mineralization protein-1 gene-transfected bone marrow mesenchymal stem cells[J]. Chinese Journal of Tissue Engineering Research, 2014, 18(23): 3633-3638.

Figures/Tables 1

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