The regulatory effect of human bone morphogenetic protein 7 gene transfer on the proliferation and differentiation of rabbit bone marrow mesenchymal stem cells.
- Author:
Dan JIN
1
;
Guo-xian PEI
;
Ke WANG
;
Kuan-hai WEI
;
Bin CHEN
;
Yu QIN
Author Information
- Publication Type:Journal Article
- MeSH: Alkaline Phosphatase; biosynthesis; Animals; Bone Marrow Cells; cytology; enzymology; Bone Morphogenetic Protein 7; Bone Morphogenetic Proteins; genetics; Cell Division; Cells, Cultured; Gene Transfer Techniques; Humans; Osteoblasts; cytology; Osteogenesis; Rabbits; Stem Cells; cytology; enzymology; Tissue Engineering; Transforming Growth Factor beta; genetics
- From: Acta Academiae Medicinae Sinicae 2003;25(1):22-25
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo detect the proliferation and differentiation of rabbit bone marrow mesenchymal stem cells (BMSc) transferred by retroviral vector carrying human bone morphogenetic protein 7 (hBMP-7) gene.
METHODShBMP-7-expressing replication-deficient retroviral vector(PT-PLNCX2-hBMP7) was reconstructed using clone technique and recombinant DNA technique. BMSc were infected with the virus granules. The protein of BMP-7 gene in transferred cells were determined by immunohistochemistry. The proliferativity of the transferred cell were assayed by methabenzthiazuron (MTT) method and flow cytometer. Alkaline phosphatase (ALP) were also detected using enzyme kinetics.
RESULTSCells transferred by PT-PLNCX2-hBMP7 expressed abundant hBMP7 protein in the cytoplasm. Positive findings were not found in those cells that were not transferred. After infected with virus there were not significant difference of cell proliferation and cell cycle between the cells transferred by hBMP-7 or not (P > 0.05). ALP activity in transferred cells were increased significantly (P < 0.01).
CONCLUSIONShBMP-7 can be transferred and stably expressed in the cultured rabbit bone marrow stem cells. Proliferation and cell cycle of the transferred cell were not affected. hBMP7 gene transfer can be used to induce differentiation of BMSc into osteoblast-like cells.
