Electransfection With Telomerase Catalytic Subunit(hTERT) Enhances Proliferative Capacity Of ADSCs.

1.Objective ADSCs offer considerable possibilities for regenerative medicine.These cells have proven to be multipotent in vitro.But the capacity of replication declined with subculturing.After 36±3 passages,these cells could no longer reach confluence.We transducted with telomerase catalytic subunit(hTERT) into ADSCs to enhance the proliferative capacity with electransfection technique and observed the effect on the proliferative capacity of ADSCs after transducted with hTERT.2.Method(1) The entire hTERT coding sequence was obtained from Genbank.Two suitable primers were designed.The hTERT RNA was derived from esophagus cancer tissue.After amplified by PCR,we connected the pcDNA3.1 plasmid and hTERT cDNA after digested the pcDNA3.1 with Hindlll.(2) The ADSCs were derived from adipose tissue from SD rats after cutting the adipose tissue into 1mm pieces and digested the tissue with collagenase l.(3) With electransfection technique,we transfected vector into ADSCs. The PCR,TRAP-PCR,and Pl-annexin V were tested to prove the expression of hTERT in ADSCs.3.ResultWe successfully transducted with telomerase catalytic subunit(hTERT) into ADSCs to enhance the prolifertative capacity convinced with PAGE and TRAP tests.The Pl-annexin V flow cytometry analyse showed the apoptosis ratio of hTERT-ADSCs was significantly lower than ADSCs.