[Studies on the transfection of umbilical endothelia with catalytic subunit of telomerase].
OBJECTIVE
To investigate the variety of proliferating ability of umbilical endothelia (UE) transfected by plasmid pBABE-HYGR-hTERT.


METHODS
UE was identified from two aspects: morphology and CD34 labeling technique. The plasmid was obtained and identified by alkali splitting and gel electrophoresis. Liposomes were used to transfect UE. RT-PCR based telomeric repeat amplification protocol (TRAP) assay was used to measure the telomerase activity of endothelia.


RESULTS
UE arranged as "cobblestone" and were positive of CD34 labeling. Endothelia transfected by pBABE-HYGR-hTERT(HC) had an raised absorbance of 0.889. The shape of growth curve of HC was similar to UE. But the absorbance of MTT test and the amount of HC were prior to UE at every measuring time and the amount of HC increased four times within 8 days (P < 0.05).


CONCLUSIONS
The transfection of pBABE-HYGRO-hTERT had greatly improved the proliferating abilities and activated the telomerase of UE.
Dai
Xiao-ming
XM
Key Laboratory of Oral Biomedical Engineering Ministry of Education, Sichuan University, Chengdu 610041, China.
Li
Long-jiang
LJ
Wen
Yu-ming
YM
Wang
Chang-mei
CM
Liu
Hua
H
Liu
Kun
K
Li
Sheng-fu
SF
D020134:Catalytic Domain / Q000235:genetics*; D002478:Cells, Cultured; D004730:Endothelium, Vascular / Q000166:cytology*; D006801:Humans; D019098:Telomerase / Q000235:genetics*; D014162:Transfection; D014471:Umbilical Veins / Q000166:cytology*
D019098:Telomerase