Specific induction and long-term maintenance of high purity ventricular cardiomyocytes from human induced pluripotent stem cells
Fig 2
Induction and maintenance of hiPSC-derived CMs with high-purity.
(A) Representative flow cytometry analysis of cTnT (and side scatter plot (SSC)) on d21 induced by combinations of Wnt signal inhibitors (XAV939 and IWP4). The percentages of cTnT+ and cTnT- cells are indicated. (B) Quantitative analysis of cTnT+ CMs on d21. Data are means ± SD. ***p < 0.001 by one-way ANOVA followed by Tukey-Kramer post hoc test (n = 4). (C) CM yield on d21. Data are means ± SD. ***p < 0.001 by one-way ANOVA followed by Tukey-Kramer post hoc test (n = 4). (D) Scheme of CM long-term maintenance protocol. CMs were re-cultured under αMEM medium with 10% FBS and Y27632, for the initial 2 days, followed by a serum-free condition (RPMI1640+B27) for 18 days. CMs underwent passage every 20 days. (E) Representative flow cytometry analysis of cTnT in 836 B3 cell line on d61, and d91. Percentages of cTnT+ cells are indicated. (F) Quantitative analysis of cTnT+ CMs in 836 B3 cell line on d61, and d91. (n = 4) (G) Representative flow cytometry analysis of cTnT in 836 B3 cell line on d201. Percentages of cTnT+ cells are indicated. (H) Representative flow cytometry analysis of cTnT in 253G1 and 1201C1 cell lines on d21, d61, and d91. Percentages of cTnT+ cells are indicated. (I) Quantitative analysis of cTnT+ CMs in 253G1 and 1201C1 cell lines on d21, d61, and d91. (253G1; n = 4, 1201C1; n = 5).