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[Immortalization of the rabbit mandibular condylar chondrocyte with SV40]
1Department of Oral Biology, College of Stomatology, Fourth Military Medical University, Xi'an 710032, China.
Objective:
To establish an immortalized cell line of the mandibular condylar chondrocyte (MCC).
Methods:
Reconstructed retroviral vector pLN/SV40 containing the large T antigen and neomycin-resistance genes were transfected into the packaging cells PA317 using lipofectin transfection method. Then the primarily cultured cells derived from mandibular condylar chondrocytes(MCCs) of New Zealand rabbits were infected with the positive retrovirus medium and monoclone was selected with G418. Specific primer of large T antigen was designed and PCR was used to detect the integrating of large T antigen gene. The positive cell clones were subcultured and their growth characteristics were compared with untransfected cells.
Results:
10 cell clones of the transduced cells showed the positive gene fragment of 1,196 bp, and 5 of them had been further subcultured for more than 30 passages. One clone of A21 had been passaged for more than 100 times in nearly one and half year, and named immortalized mandibular condylar chondrocyte (IMCC). IMCCs were polygonal-shaped, similar to MCCs; the population doubling time of IMCC and MCC were 30.86 h and 78.95 h respectively. Subculture, freezing and recovering had no effect on cellular shape and proliferation of IMCC.
Conclusions:
An immortalized cell cline derived from the rabbit MCC has been successfully established.