Corning® 500 mL RPMI 1640

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Publication protocol

Cardiac differentiation TIMING ~15–20 days. Once the hiPSCs have reached confluency (Day 0), change the media to RPMI 1640 medium containing B27 supplement minus insulin (RPMI/B27 -I) and 10 μM CHIR and incubate at 37 °C and 5% CO2 for 24h. Washing plates between this and the following media changes is not required. On Day 1, change the media to RPMI/B27-I (no small molecule) and incubate (37 °C and 5% CO2) for 24 h. On Day 2, change the media to RPMI/B27-I containing 5 μM of IWP4 and incubate at 37 °C and 5% CO2 for 48h. On Day 4, change the media back to RPMI/B27-I (no small molecule) for 2 additional days. On Day 6, change the media to RPMI 1640 containing B27 complete supplement (RPMI/B27+C). Repeat the media change with RPMI/B27+C the following day (Day 7). From here on, change the media every two days until Day 20. The formation of 3D structures should start from Day 1 after CHIR treatment, and beating cardiac microchambers should be visible around Day 10.

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