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Traditionally, two major approaches have been used to immortalize primary cells, including the serial passages of primary cells until they overcome their growth-crisis stage and the transformation of primary cells by overexpression of one or more oncogenes. The serial passage has been used to establish the Balb/3T3 cell line [37]. However, most of these established cell lines were hypotetraploid [37]. Furthermore, it is unclear if these transformed cells retain multi-lineage differentiation potential. Numerous studies have been recently carried out to induce the pluripotency of fibroblasts and cardiomyogenic progenitors through reprogramming stem cell differentiation capability with a set of defined factors, such as Oct3/4, Sox9, Klf4, and c-Myc [38-42] or small molecules [43]. Our results have demonstrated that the reversible immortalization strategy using SV40 large T antigen is an efficient approach to establishing long-term cell culture of primary CPs for basic and translational research..

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