ARTICOLI ORIGINALI   

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

Copyright © 2014 EDIZIONI MINERVA MEDICA

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.