title: [Regulation of human bone marrow mesenchymal stem cells osteogenic and adipogenic differentiations by Wnt10b adenoviral vector in vitro].
abstract: OBJECTIVE
  To investigate the regulation of human bone marrow mesenchymal stem cells (hBMSCs) osteogenic and adipogenic differentiations mediated by Wnt10b adenoviral vector in vitro.
  
  
  METHODS
  The hBMSCs from ilial bone tissue in adults at passage 4 were infected by Wnt10b gene expression adenoviral vector (group A), Wnt10b-shRNA adenoviral vector (group B), and empty vector (group C), and non-transfected hBMSCs served as the blank control group. Then the cells were cultured separately in the circumstance of osteogenic induction, adipogenic induction, and non-induction. The alkaline phosphatase (ALP) staining, alizarin red staining, and oil red O staining were used to detect the osteogenic and adipogenic differentiations; real-time fluorescent quantitative PCR and Western blot were used to analyze the expressions of osteoblast and adipocyte genes and proteins.
  
  
  RESULTS
  The results of ALP staining were positive after osteogenic induction, group A showed strong staining, and group B showed the weakest staining. The results of alizarin red staining showed that there were a lot of patchy confluent brown mineralized nodules in group A; a few punctate brown mineralized nodules were seen in group B; and many punctuate brown mineralized nodules were found in groups C and D. The results of oil red O staining showed strong staining in groups B, C, and D after adipogenic induction, especially in group B; scattered or small clustered staining was observed in group A. The expressions of osteoblast genes and proteins were significantly higher in group A than groups B, C, and D, and in groups C and D than group B by real-time fluorescent quantitative PCR and Western blot test; however, the expressions of adipocyte genes and proteins showed a contrary tendency.
  
  
  CONCLUSIONS
  The high level expression of Wnt10b can enhance osteogenic differentiation of hBMSCs, and the low level expression of Wnt10b can increase adipogenic differentiation of hBMSCs.
authors.affiliation: Department of Orthopaedics, West China Hospital, Sichuan University, Chengdu Sichuan, P.R. China.
authors.forename: Yong
authors.initials: Y
authors.lastname: Zhou
authors.forename: Chong
authors.initials: C
authors.lastname: Zou
authors.forename: Li
authors.initials: L
authors.lastname: Min
authors.forename: Rui
authors.initials: R
authors.lastname: Shi
authors.forename: Wenli
authors.initials: W
authors.lastname: Zhang
authors.forename: Yi
authors.initials: Y
authors.lastname: Luo
authors.forename: Jing
authors.initials: J
authors.lastname: Peng
authors.forename: Hong
authors.initials: H
authors.lastname: Duan
authors.forename: Chongqi
authors.initials: C
authors.lastname: Tu
authors.forename: Hui
authors.initials: H
authors.lastname: Zhang
mesh_terms: D000256:Adenoviridae; D017667:Adipocytes; D000328:Adult; D015153:Blotting, Western; D001854:Bone Marrow Cells / Q000378:metabolism*; D001842:Bone and Bones; D002454:Cell Differentiation; D002478:Cells, Cultured; D015870:Gene Expression; D005822:Genetic Vectors; D006412:Hematopoietic Stem Cells; D006801:Humans; D066298:In Vitro Techniques; D010006:Osteoblasts / Q000378:metabolism*; D010012:Osteogenesis / Q000235:genetics*; D011518:Proto-Oncogene Proteins / Q000378:metabolism*; D034741:RNA, Small Interfering; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014162:Transfection; D051153:Wnt Proteins / Q000378:metabolism*
chemical_list: D011518:Proto-Oncogene Proteins; D034741:RNA, Small Interfering; C491119:WNT10B protein, human; D051153:Wnt Proteins