Osteogenic differentiation of adipose-derived stem cells transfected by lentivirus vector carrying LIM mineralization protein-1 and hypoxia-inducible factor-1alpha

doi:10.3969/j.issn.2095-4344.2015.32.011

Abstract

Abstract:

BACKGROUND: LIM mineralization protein-1 (LMP-1) and hypoxia-inducible factor-1α (HIF-1α) as intracellular proteins can induce osteogenic differentiation and promote angiogenesis, respectively. Therefore, their combination is of great significance for effectively inducing the osteogenic differentiation of adipose-derived stem cells.
OBJECTIVE: To study the osteogenic differentiation of adipose-derived stem cells transfected by lentivirus vector carrying LMP-1 and HIF-1α.
METHODS: Reverse transcription-PCR technology was employed to clone LMP-1 and HIF-1α genes, and the genes were cloned to lentivirus vectors pLVX-EF1α-DsRed-Hyg and pLVX-EF1α-IRES2-AcGFP1 to construct main lentiviral vectors pLVX-EF1α-DsRed-Hyg-RLMP-1 and pLVX-EF1α-IRES2-AcGFP1-RHIF-1α. Then, Lenti-X 293T cells were transfected with main vectors pLVX-EF1α-DsRed-Hyg-RLMP-1 and pLVX-EF1α-IRES2-AcGFP1-RHIF-1α, packaging plasmid and coated plasmid. After that, lentiviral vectors were packaged to transfect adipose-derived stem cells from rats that were obtained by tissue explants culture and enzyme digestion methods. At 3, 7, 14 days after transfection, reverse transcription-PCR technology was adopted to detect the expression of osteogeic genes, such as bone morphogenetic protein 2, Runx-2, alkaline phosphatase, osteocalcin as well as to detect the expression of vascular endothelial growth factor.
RESULTS AND CONCLUSION: Lentiviral vectors pLVX-EF1α-DsRed-Hyg-RLMP-1 and pLVX-EF1α-IRES2-AcGFP1-RHIF-1α were effectively transfected into adipose-derived stem cells. Reverse transcription-PCR results showed that from the 7th day to the 14th day after lentivirus transfection, bone morphogenetic protein 2, Runx-2, alkaline phosphatase and osteocalcin all over-expressed. These findings indicate that the combination of LMP-1 and HIF-1α can enhance the osteogenic activity of adipose-derived stem cells.

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

Key words: Transfection, Cell Differentiation, Viruses

CLC Number:

R394.2

Pan Wei-min, Liu Min, Yang Jian-chang, Duan Chun-guang, Huang Yue. Osteogenic differentiation of adipose-derived stem cells transfected by lentivirus vector carrying LIM mineralization protein-1 and hypoxia-inducible factor-1alpha[J]. Chinese Journal of Tissue Engineering Research, 2015, 19(32): 5140-5147.

Figures/Tables 4

References

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