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Indexed/Abstracted in: Current Contents/Clinical Medicine, EMBASE, PubMed/MEDLINE, Science Citation Index Expanded (SciSearch), Scopus
Impact Factor 1,236
Online ISSN 1827-1669
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.