[The regulatory effect of human bone morphogenetic protein 7 gene transfer on the proliferation and differentiation of rabbit bone marrow mesenchymal stem cells].
OBJECTIVE
To 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.


METHODS
hBMP-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.


RESULTS
Cells 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).


CONCLUSIONS
hBMP-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.
Jin
Dan
D
Department of Orthopedics and Traumatology, Nanfang Hospital, First Military Medical University, Guangzhou 510515, China. nfjindan@263.net
Pei
Guo-xian
GX
Wang
Ke
K
Wei
Kuan-hai
KH
Chen
Bin
B
Qin
Yu
Y
D000469:Alkaline Phosphatase / Q000096:biosynthesis; D000818:Animals; D001854:Bone Marrow Cells / Q000166:cytology / Q000201:enzymology; D055419:Bone Morphogenetic Protein 7; D019485:Bone Morphogenetic Proteins / Q000235:genetics*; D002455:Cell Division; D002478:Cells, Cultured; D018014:Gene Transfer Techniques; D006801:Humans; D010006:Osteoblasts / Q000166:cytology*; D010012:Osteogenesis; D011817:Rabbits; D013234:Stem Cells / Q000166:cytology* / Q000201:enzymology; D023822:Tissue Engineering; D016212:Transforming Growth Factor beta / Q000235:genetics*
C525819:BMP7 protein, human; D055419:Bone Morphogenetic Protein 7; D019485:Bone Morphogenetic Proteins; D016212:Transforming Growth Factor beta; D000469:Alkaline Phosphatase