Extension of Cell Life Span Using Exogenous Telomerase

Normal human somatic cells undergo limited cell division cycles and enter irreversible replication arrest called senescence. Cellular senescence of many human cell types is regulated by the length and status of telomeric sequences, which is shortened after each round of DNA replication. Telomeres can be rejuvenated by telomerase, an enzyme which carries out de novo synthesis of telomeric DNA. Telomerase is a ribonucleoprotein complex composed minimally of telomere reverse transcriptase gene (hTERT ) and RNA template (hTR ), and its enzyme activity in cells is primarily limited by the level of hTERT expression. Therefore, telomerase activity in cells can be reconstituted by overexpression of hTERT , frequently resulting in extension of replicative life span or immortalization. It is well established that the effect of telomerase reconstitution on cellular life span is clearly cell typedependent because telomere shortening is not the only limiting factor of cellular life span. However, telomerase activity appears to be a requirement for cellular immortalization, irrespective of the cell types. In this article, we discuss the detailed methods to extend the in vitro replicative life span of primary human cells by ectopic expression of hTERT .