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Figure 4 | Stem Cell Research & Therapy

Figure 4

From: Excision of viral reprogramming cassettes by Cre protein transduction enables rapid, robust and efficient derivation of transgene-free human induced pluripotent stem cells

Figure 4

Improved cardiac differentiation potential of transgene-free induced pluripotent stem cells. (A) Genomic polymerase chain reaction (PCR) for the confirmation of transgene deletion. fl-ARiPS clone 1 was treated with 2 μM TAT-Cre for 5 hours and PCR was performed using primers against WPRE and β-actin (BA). (B) Cardiomyocyte differentiation of iPSCs. fl-ARiPS clone 1 and its TAT-Cre-treated polyclonal cell progeny were differentiated towards cardiomyocyte lineage using cardio-inductive medium. Cells were stained with α-actinin antibody at day 15. Scale bar: 40 μm. (C) Flow cytometry analysis of cardiac-specific troponin T (cTNT) staining at day 15 of cardiac differentiation. Cardiac differentiation showed an increase from 37 to 56% cTNT-positive cardiomyocytes in the case of TAT-Cre-treated cell populations. The experiment was repeated twice with similar results.

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