The effect of chTERT gene transfer on the feeder layer cells of chicken primodial germ cells

Hsiao Yun Kuo and Lih-Ren Chen

　　Telomere-dependent replicative senescence is one of the mechanisms that limit the doubling number of normal cells. By overexpression of telomerase, cells of various origins have been successfully immortalized without changing the phenotype. The telomerase have three main parts: including telomerase RNA (hTR), TPI/TLPI and telomerase reverse transcriptase (TERT). The TERT, which is the catalytic component of telomerase, and the primary function of telomerase is to maintain telomere length in the cells. In the previous study, the major problem was the feeder layer of PGC unable to live long enough to support the PGC growth in vitro. Feeder cells used in this study was stromal cells separated from the primary germline during collecting poultry PGC. Hence, the purpose of this study is to construct the vector containing TERT, and transfer into stromal cells by electroporation and lentivirus. The results showed that although stromal cells can be transfected with TERT gene carriers and then extended the lifespan, but still have the problem of poor transfection efficiency, low cell survival rate and low cell division rate. This would also affect subsequent conduct analysis of telomere length and telomerase activity. The results still have follow-up efforts to achieve improvement of long-term culture of PGC in vitro. We would continue to adopt viral transfection and telomere length and activity analysis methods which established in this trial to extend the lifespan of stromal cells, and the efficiency of providing stable feeders in the future.