PMID- 24501901
OWN - NLM
STAT- MEDLINE
DCOM- 20140702
LR  - 20181202
IS  - 1002-1892 (Print)
IS  - 1002-1892 (Linking)
VI  - 27
IP  - 11
DP  - 2013 Nov
TI  - [Osteodifferentiation of bone marrow mesenchymal stem cells after transfected by 
      lentiviral vector mediated bone morphogenetic protein 2].
PG  - 1380-5
AB  - OBJECTIVE: To construct recombinant lentiviral vectors of porcine bone 
      morphogenetic protein 2 (BMP-2) gene and to detect BMP-2 gene activity and bone 
      marrow mesenchymal stem cells (BMSCs) osteogenetic differentiation so as to lay a 
      foundation of the further study of osteochondral tissue engineering. METHODS: 
      BMSCs were isolated from bone marrow of 2-month-old Bama miniature porcines 
      (weighing, 15 kg), and the 2nd generation of BMSCs were harvested for 
      experiments. The porcine BMP-2 gene lentiviral vector was constructed by 
      recombinant DNA technology and was used to transfect BMSCs at multiplicity of 
      infection (MOI) of 10, 25, 50, 100, and 200, then the optimal value of MOI was 
      determined by fluorescent microscope and inverted phase contrast microscope. 
      BMSCs transfected by BMP-2 recombinant lentiviral vectors served as experimental 
      group (BMP-2 vector group); BMSCs transfected by empty vector (empty vector 
      group), and non-transfected BMSCs (non-transfection group) were used as control 
      groups. RT-PCR, immunohistochemistry staining, and Western blot were performed to 
      detect the expressions of BMP-2 mRNA and protein. Then the BMSCs osteogenesis was 
      detected by alkaline phosphatase (ALP) staining, ALP activities, and Alizarin red 
      staining. RESULTS: The recombinant lentiviral vectors of porcine BMP-2 gene was 
      successfully constructed and identified by RT-PCR and gene sequencing, and BMSCs 
      were successfully transfected by BMP-2 recombinant lentiviral vectors. Green 
      fluorescent protein could be seen in the transfected BMSCs, especially at MOI of 
      100 with best expression. The immunohistochemistry staining and Western blot 
      showed that BMSCs transfected by BMP-2 recombinant lentiviral vectors could 
      express BMP-2 protein continuously and stably at a high level. After cultivation 
      of 2 weeks, the expression of ALP and the form of calcium nodules were observed. 
      CONCLUSION: The porcine BMP-2 gene lentiviral vector is successfully constructed 
      and transfected into the BMSCs, which can express BMP-2 gene and protein 
      continuously and stably at a high level and induce BMSCs differentiation into 
      osteoblasts.
FAU - Xu, Shenggui
AU  - Xu S
AD  - Department of Orthopedics, Mindong Hospital Affiliated to Fujian Medical 
      University, Ningde Fujian, 355000, P.R.China.
FAU - Lin, Jianhua
AU  - Lin J
AD  - Department of Orthopedics, the First Affiliated Hospital of Fujian Medical 
      University.
FAU - Liu, Weinan
AU  - Liu W
AD  - Department of Orthopedics, the Affiliated People's Hospital of Fujian University 
      of Traditional Chinese Medicine.
FAU - Wu, Chaoyang
AU  - Wu C
AD  - Department of Orthopedics, the First Affiliated Hospital of Fujian Medical 
      University.
LA  - chi
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - China
TA  - Zhongguo Xiu Fu Chong Jian Wai Ke Za Zhi
JT  - Zhongguo xiu fu chong jian wai ke za zhi = Zhongguo xiufu chongjian waike zazhi = 
      Chinese journal of reparative and reconstructive surgery
JID - 9425194
RN  - 0 (Bone Morphogenetic Protein 2)
RN  - 0 (RNA, Messenger)
RN  - 0 (Recombinant Proteins)
RN  - 0 (Transforming Growth Factor beta)
RN  - 0 (recombinant human bone morphogenetic protein-2)
RN  - EC 3.1.3.1 (Alkaline Phosphatase)
SB  - IM
MH  - Adenoviridae/*genetics
MH  - Alkaline Phosphatase/metabolism
MH  - Animals
MH  - Bone Marrow Cells/cytology/metabolism
MH  - Bone Morphogenetic Protein 2/*genetics/metabolism
MH  - *Cell Differentiation
MH  - Cell Proliferation
MH  - Cells, Cultured
MH  - Female
MH  - Gene Expression
MH  - Genetic Therapy
MH  - Genetic Vectors/*genetics
MH  - Male
MH  - Mesenchymal Stem Cells/*cytology/metabolism
MH  - *Osteogenesis
MH  - RNA, Messenger/genetics/metabolism
MH  - Recombinant Proteins/genetics/metabolism
MH  - Swine
MH  - Tissue Engineering/methods
MH  - Transfection
MH  - Transforming Growth Factor beta/*genetics/metabolism
EDAT- 2014/02/08 06:00
MHDA- 2014/07/06 06:00
CRDT- 2014/02/08 06:00
PHST- 2014/02/08 06:00 [entrez]
PHST- 2014/02/08 06:00 [pubmed]
PHST- 2014/07/06 06:00 [medline]
PST - ppublish
SO  - Zhongguo Xiu Fu Chong Jian Wai Ke Za Zhi. 2013 Nov;27(11):1380-5.