Differentiation

In the Foo lab, we are continuously generating hES-derived cardiomyocytes as an in-vitro model of human heart development and disease. This process is called differentiation.

The protocol in our Lab was optimised and improved from a previously published protocol by Lian et al (Lian et al. 2012). Small molecule CHIR99021 (GSK3 inhibitor) is added on day 0 and left for 24 hours followed by media change. On day 3, IWP2 (WNT inhibitor) is added using 50/50 mix of new fresh medium and conditioned medium collected from each well and left for 48 hours. Culture medium from day 0 until day 7 is RPMI1640 plus B-27 serum-free supplement without insulin. From day 7 and onwards, RPMI1640 with B-27 serum free supplement (with insulin) is used and changed every 2-3 days. With 80-95% differentiation efficiency, cardiomyocytes start to beat from day 14.