Systematic Following of Telomerase during MRC5 Population Dubbling and Cell Senescence

Abstract

MRC5 are well-characterized human diploid fibroblast cell line approved for vaccine production. We have adapted measles, Rubella and Polio viruses for propagation in MRC5 cell lines. In this study, telomerase activity during different passages of this cell detected and traced. Stocks of MRC5 cells from passage 15 to passage 38 were monitored. Trace amount of telomerase activity detected in these cells using TRAP-PCR with gel based detection. A decreasing order of telomerase activity observed with the cell doubling number. In order to compare in more details the telomerase activity during cell passage and replicative senescence, it was reassayed with an ultra sensitive method of TRAP-PCR ELISA based method. While in gel-based method trace amount of telomerase activity decreasing with increasing passage number of cells were detected, in PCR-ELISA very lower detectable telomerase activity were observed. We think this could be because of low processivity of this enzyme.KeywordsTelomeraseMCR5TRAP-PCR