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. 2021 Jun 4;2(6):100298. doi: 10.1016/j.xcrm.2021.100298

Figure 1.

Figure 1

The modified replating method improved the differentiation and maturation of hiPSC skeletal myotubes

(A) A schematic diagram of the modified replating method. iPSCs (409B2, 409B2 ex45KO, DMDΔ44-ctrl, and DMDΔ44) were myogenically pre-differentiated in Primate ES Cell Medium (PECM) media in the presence of doxycycline. Cells were replated on day 4 in the presence of lower doses of Y-27632 and the absence of doxycycline in 2% horse serum media. The doxycycline-induced MyoD overexpression induced myogenic differentiation at day 6 for 4 days.

(B) Bright-field images of the time course of myogenic differentiation of 409B2 and DMDΔ44 lines. Scale bar represents 200 μm.

(C) Immunocytochemical analysis of pan-MHC and DYS1 in differentiated myotubes of 409B2 and DMDΔ44 lines on day 14. Scale bar represents 20 μm.

(D) Fusion index analysis of 409B2, 409B2 ex45KO, DMDΔ44-ctrl, and DMDΔ44 myotubes differentiated by previous and modified replating methods. Data represent the mean ± SD and were analyzed by an unpaired t test from six biological replicates. ∗p < 0.05.