Journal: International Journal of Molecular Sciences

Article Title: Differential Sarcomere and Electrophysiological Maturation of Human iPSC-Derived Cardiac Myocytes in Monolayer vs. Aggregation-Based Differentiation Protocols

doi: 10.3390/ijms18061173

Figure Lengend Snippet: High content cell imaging to analyze the cell size, nucleation and morphology using the cTnT and Dapi staining. ( A ) Design of the automated cellular imaging high content analysis; ( B ) Composite image of the detection of cTnT+ and Dapi+ cells. Detection setting for total, round and long cells ( C ) Cell surface comparison at day 27 ± 2. N > 1500 cells from at least three different differentiations with three different iPSC clones, **** p < 0.0001; ( D ) Distribution of cell surface area in generated cTNT+ cells from the 2D-IWR1 (green) and 2D-IWP2 (blue); ( E ) Frequency of number of nuclei in the total cells for each protocol ( F ) Distribution of round and long cells obtained with each protocol. ** p < 0.01.

Article Snippet: The cardiac progenitors were induced on day 3 with 2D differentiation medium supplemented with either a Wnt inhibitor IWR1 (Inhibitor of Wnt Response, 2.5 μM) or IWP2 (Inhibitor of Wnt Production, 5 μM) (Tocris, Bristol, USA) for 2 days.

Techniques: Imaging, Staining, High Content Screening, Comparison, Clone Assay, Generated

Journal: International Journal of Molecular Sciences

Article Title: Differential Sarcomere and Electrophysiological Maturation of Human iPSC-Derived Cardiac Myocytes in Monolayer vs. Aggregation-Based Differentiation Protocols

doi: 10.3390/ijms18061173

Figure Lengend Snippet: Quantification of sarcomere mRNA and proteins. ( A ) The mRNA expression of cTni and ssTni was quantified by quantitative RT PCR. ( n = 8 to 12 from four iPSC clones, p < 0.05 Kruskal-Wallis and Dunn’s Multiple Comparison Test, hVentriCM was excluded from statistical analysis); ( B ) Typical western blot of cTNi and ssTNi proteins; ( C ) Quantification of the ratio of cTNi/GAPDH on ssTNi/GAPDH proteins in hiPSC-CM generated with the 2D-IWR1 vs. 2D-IWP2 protocols. Quantifications from 4 differentiations with 4 different iPS clones, * p < 0.05, Mann–Whitney test.

Article Snippet: The cardiac progenitors were induced on day 3 with 2D differentiation medium supplemented with either a Wnt inhibitor IWR1 (Inhibitor of Wnt Response, 2.5 μM) or IWP2 (Inhibitor of Wnt Production, 5 μM) (Tocris, Bristol, USA) for 2 days.

Techniques: Expressing, Quantitative RT-PCR, Clone Assay, Comparison, Western Blot, Generated, MANN-WHITNEY

Journal: International Journal of Molecular Sciences

Article Title: Differential Sarcomere and Electrophysiological Maturation of Human iPSC-Derived Cardiac Myocytes in Monolayer vs. Aggregation-Based Differentiation Protocols

doi: 10.3390/ijms18061173

Figure Lengend Snippet: Main electrophysiological parameters. (*) indicates significances levels of p < 0.05 and (**) p < 0.01 3D vs. 2D-IWR1. ( $ ) indicates significances levels of p < 0.05 and ( $$ ) p < 0.01 3D vs. 2D-IWP2 ( £ ) indicates significances levels of p < 0.05 and ( ££ ) p < 0.01 2D-IWR1 vs. 2D-IWP2.

Article Snippet: The cardiac progenitors were induced on day 3 with 2D differentiation medium supplemented with either a Wnt inhibitor IWR1 (Inhibitor of Wnt Response, 2.5 μM) or IWP2 (Inhibitor of Wnt Production, 5 μM) (Tocris, Bristol, USA) for 2 days.

Techniques: