[Reconstitution of telomerase activity in human embryonic tendon cells transfected by ptsA58H plasmid].
OBJECTIVE
To investigate the feasibility of life span extension of transformed human embryonic tendon cells (THETC) by reconstitution of the telomerase activity.


METHODS
THETC were transfected by pGRN145 plasmid containing the human telomerase reverse transcriptase (hTERT) cNDA in vitro by molecular cloning technique. The biological characteristics of transfected cells were detected and compared by morphological observation, plate cloning efficiency, soft agar culture, growth curve of cells cultured in different conditions, immunohistochemistry, telomerase activity assay by telomeric repeat amplification protocol (TRAP).


RESULTS
The THETC transfected by pGRN145 plasmid (telT) could express the telomerase activity with extension of life span. The telT maintained the original characteristics of temperature-dependant and serum-dependant, as well as secretion of type I collagen normally and without tendency of malignant transformation.


CONCLUSIONS
The life span of THETC can be prolonged by reconstitution of telomerase activity, which provides the novel experimental methods to establish the standard cells line.
Xie
Hui-qi
HQ
Institute of Reparative and Reconstructive Surgery, West China Hospital, Sichuan University, Chengdu, Sichuan 610041, China.
Qu
Yi
Y
Li
Xiu-qun
XQ
Qin
Ting-wu
TW
Yang
Zhi-ming
ZM
D002460:Cell Line; D002470:Cell Survival; D004622:Embryo, Mammalian; D006801:Humans; D010957:Plasmids / Q000235:genetics; D012194:RNA-Directed DNA Polymerase; D019098:Telomerase / Q000235:genetics* / Q000378:metabolism; D013710:Tendons / Q000166:cytology* / Q000201:enzymology*; D014162:Transfection
D012194:RNA-Directed DNA Polymerase; D019098:Telomerase