Research on bone marrow derived-mesenchymal stem cells and human umbilical cord blood derived-mesenchymal stem cells modified with the gene of human bone morphogenetic protein-2

doi:10.3877/cma.j.issn.2096-0263.2019.02.004

Abstract

Abstract:

Objective

To study the modification of bone marrow and human umbilical cord blood mesenchymal stem cells with hBMP-2 gene and compare the modification results.

Methods

Bone marrow and human umbilical cord blood mesenchymal stem cells were separated and cultured by density gradient centrifugation and adherent culture. The recombinant hBMP-2 plasmid was transfected into bone marrow and human umbilical cord blood mesenchymal stem cells by high-performance non-liposome reagent X-treme GENE. The fluorescence intensity was measured by inverted fluorescence microscopy and the transfection efficiency was calculated. The hBMP was detected by qPCR. The expression of-2 and chondronectin in two kinds of cells was detected by immunohistochemistry.

Results

Bone marrow and human umbilical cord blood mesenchymal stem cells were successfully isolated and cultured. The two cells had different growth characteristics. The recombinant hBMP-2 plasmid mediated by high-performance non-liposome reagent was successfully transfected into bone marrow and human umbilical cord blood mesenchymal stem cells. The efficiency of transfection of bone marrow mesenchymal stem cells (18.44±5.94)% was lower than that of human umbilical cord blood mesenchymal stem cells (27.74±7.59)%, and the difference was statistically significant (t=3.027, P<0.05). The expression of hBMP-2, cartilage connectin and collagen type II were detected in both transfected cells.

Conclusion

hBMP-2 gene can effectively modify bone marrow and human umbilical cord blood mesenchymal stem cells and promote chondrocyte differentiation.

Key words: BMP2 protein, human, Mesenchymal stem cells, Liposomes

Jie Zhang, Hui Huang, Junguo Wang, Yi Sun, Haining Zhang, Yingzhen Wang. Research on bone marrow derived-mesenchymal stem cells and human umbilical cord blood derived-mesenchymal stem cells modified with the gene of human bone morphogenetic protein-2[J]. Chinese Journal of Geriatric Orthopaedics and Rehabilitation(Electronic Edition), 2019, 05(02): 82-86.

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