[Immortalization of human osteoblast by transferring hTERT gene].
OBJECTIVE
To establish an immortalized osteoblast cell line used for basal and clinical research of orthopaedics.


METHODS
Human marrow stromal cells (hMSCs) had been inducted into osteoblasts directionally, and human telomerase reverse transcriptase (hTERT) cDNA was transferred into the osteoblasts by retroviral vector pLPChTERT. The resultant stable clones reproduced successively and the expression of hTERT as well as the osteogenesis characteristics of different eras were identified.


RESULTS
hTERT gene has been transferred into human osteoblasts successfully. The transformed cells expressed telomerase activity and divided vigorously. p62 has been obtained so far. The expression of bone specific Alkaline phosphatase, Collagen type I and Osteopontin of p25, p55 were checked and it is proved that the immortalized cell line preserved typical osteogenesis characteristics.


CONCLUSIONS
Osteoblasts can be immortalized by transferring exogenous hTERT gene to reconstitute telomerase activity, and the immortalized cell reserved osteogenesis characteristics.
Yin
Xiao-xue
XX
Department of Orthopaedics, Third Hospital, Peking University, Beijing 100083, China.
Chen
Zhong-qiang
ZQ
Guo
Zhao-qing
ZQ
Dang
Geng-ding
GD
Ma
Qing-jun
QJ
Wang
Shen-wu
SW
D001854:Bone Marrow Cells / Q000166:cytology; D002461:Cell Line, Transformed; D002478:Cells, Cultured; D016922:Cellular Senescence / Q000235:genetics; D018076:DNA, Complementary / Q000235:genetics; D004268:DNA-Binding Proteins; D018014:Gene Transfer Techniques; D005822:Genetic Vectors / Q000235:genetics; D006801:Humans; D010006:Osteoblasts / Q000166:cytology*; D017154:Stromal Cells / Q000166:cytology; D019098:Telomerase / Q000096:biosynthesis / Q000235:genetics*; D014162:Transfection
D018076:DNA, Complementary; D004268:DNA-Binding Proteins; D019098:Telomerase