title: Transfection of hBMP-2 into mesenchymal stem cells derived from human umbilical cord blood and bone marrow induces cell differentiation into chondrocytes.
abstract: OBJECTIVE
  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.
  
  
  CONCLUSIONS
  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.
authors: Qilu Hospital of Shandong University Jinan, China2Department of Orthopedics Qingdao Hiser Medical Center, Qingdao, China3Department of Joint Surgery Affiliated Hospital of Qingdao University Qingdao, China - orthopedist_qd@163.com.
authors: J
authors: J
authors: Wang
authors: J
authors: J
authors: Li
authors: N
authors: N
authors: Deng
authors: X
authors: X
authors: Zhao
authors: Y
authors: Y
authors: Liu
authors: X
authors: X
authors: Wang
authors: H
authors: H
authors: Zhang
mesh_terms: D055396:Bone Morphogenetic Protein 2 / Q000008:administration & dosage / Q000032:analysis / Q000235:genetics / Q000502:physiology*; D018929:Cell Culture Techniques; D002454:Cell Differentiation / Q000235:genetics / Q000502:physiology*; D019902:Chondrocytes / Q000166:cytology*; D024043:Collagen Type II / Q000032:analysis / Q000378:metabolism; D005312:Fetal Blood / Q000166:cytology; D049452:Green Fluorescent Proteins / Q000008:administration & dosage; D006801:Humans; D008081:Liposomes; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000378:metabolism; D039902:Multipotent Stem Cells / Q000166:cytology; D010957:Plasmids; D012333:RNA, Messenger / Q000378:metabolism; D013997:Time Factors; D014162:Transfection / Q000379:methods
chemical_list: C525718:BMP2 protein, human; D055396:Bone Morphogenetic Protein 2; D024043:Collagen Type II; D008081:Liposomes; D012333:RNA, Messenger; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins