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.
Result: 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.
Conclusion: Osteoblasts can be immortalized by transferring exogenous hTERT gene to reconstitute telomerase activity, and the immortalized cell reserved osteogenesis characteristics.