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Minerva Medica 2014 Agosto;105(4):283-8

lingua: Inglese

Transfection of hBMP-2 into mesenchymal stem cells derived from human umbilical cord blood and bone marrow induces cell differentiation into chondrocytes

Wang J. 1, 2, Li J. 1, Deng N. 2, Zhao X. 2, Liu Y. 2, Wang X. 2, Zhang H. 3

1 Qilu Hospital of Shandong University, Jinan, China;
2 Department of Orthopedics, Qingdao Hiser Medical Center, Qingdao, China;
3 Department of Joint Surgery, Affiliated Hospital of Qingdao University, Qingdao, China


AIM: Objective of the study was to compare the liposome-mediated transfection efficiency of the recombinant plasmid pIRES2-EGFP-hBMP-2 into mesenchymal stem cells (MSCs) derived from human umbilical cord blood (UCB-MSCs) and bone marrow (BM-MSCs).
METHODS: UCB-MSCs and BM-MSCs were isolated using density gradient centrifugation followed by adherent cultures. The plasmid (pIRES2-EGFP-hBMP-2) was transfected into MSCs by using X-treme GENE transfection kit. Successful transfection was determined by both the protein expression of EGFP observed by fluorescence microscopy and the mRNA expression of human bone morphogenetic protein-2 (hBMP-2) detected using RT-PCR. Cell surface marker and morphological changes were examined by immunohistochemistry (IHC) after two weeks of transfection.
RESULTS: Both UCB-MSCs and BM-MSCs were isolated and expanded successfully for multiple passages and demonstrated slight growth and morphological differences. The recombinant plasmid (pIRES2-EGFP-hBMP-2) was successfully transfected into both UCB-MSCs and BM-MSCs with an efficiency of 27.7±7.6% and 18.4±5.9%, respectively. The mRNA expression of hBMP-2 correlated with increased staining for collagen type II in the transfected cells.
CONCLUSION: Transfection of hBMP-2 in UCB-MSCs and BM-MSCs increases the expression of collagen type II. These results indicate that increased BMP-2 levels induce multipotent stem cell differentiation into chondrocytes.

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