Abstract
Integrating viruses represent robust tools for cellular reprogramming; however, the presence of viral transgenes in induced pluripotent stem cells (iPSCs) is deleterious because it holds the risk of insertional mutagenesis leading to malignant transformation. Here, we combine the robustness of lentiviral reprogramming with the efficacy of Cre recombinase protein transduction to derive iPSCs devoid of transgenes. By genome-wide analysis and targeted differentiation towards the cardiomyocyte lineage, we show that transgene-free iPSCs are superior to iPSCs before Cre transduction. Our study provides a simple, rapid and robust protocol for the generation of clinical-grade iPSCs suitable for disease modeling, tissue engineering and cell replacement therapies.
Highlights
Initial reports for generating induced pluripotent stem cells involved retroviruses as a mode of transgene delivery [1]
Retroviruses are capable of reprogramming somatic cells to induced pluripotent stem cells (iPSCs), the clinical applicability of such iPSCs is limited due to the integrated transgenes carrying the risk of insertional mutagenesis [2] and tumor formation [3]
[8], microRNAs [9], Sendai virus [10] as well as protein transduction [11,12,13]. iPSCs generated by these methods contain minimal or no genetic modifications and are generally more suitable for clinical applications than cells derived by virus-based protocols
Summary
Initial reports for generating induced pluripotent stem cells (iPSCs) involved retroviruses as a mode of transgene delivery [1]. Retroviruses are capable of reprogramming somatic cells to iPSCs, the clinical applicability of such iPSCs is limited due to the integrated transgenes carrying the risk of insertional mutagenesis [2] and tumor formation [3]. Continuous expression of transgenes in iPSCs negatively affects pluripotency [4] and limits their differentiation potential [5]. These effects have been shown by the inability to yield live chimeric mice and the diminished endodermal differentiation of iPSCs carrying transgenes [5]. IPSCs generated by these methods contain minimal or no genetic modifications and are generally more suitable for clinical applications than cells derived by virus-based protocols. In terms of robustness and efficacy, the retroviral and lentiviral system still represents the method of choice for iPSC derivation [16]
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