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. 2016 Aug 15;25(16):1243–1248. doi: 10.1089/scd.2016.0035

FIG. 1.

FIG. 1.

Generation of immortalized fibroblasts by PB-mediated gene transfer. (A) Structure of the two plasmid constructs for immortalization. pT1 contains hTERT, and pT2 contains hTERT, SV40LT, and EGFP; both have puro-TK for selection. (B) Colony morphology of fibroblasts reversibly immortalized with pT1 and pT2. (C) Growth curves of parental fibroblasts and cells immortalized with pT1 and pT2. Parental fibroblasts grew slowly and became senescent at passage 10, while pT1- and pT2-immortalized fibroblasts continued to proliferate, the latter at twice the rate of the former. (D) Number of immortalized fibroblast cell lines derived from pT1 and pT2 transfection from day 20 to 60 of culture. These cell lines grew exponentially for several months. EGFP, enhanced green fluorescent protein; hTERT, human telomerase reverse transcriptase; PB, piggyBac.