Mitochondrial Maturation in Human Pluripotent Stem Cell Derived Cardiomyocytes

Figure 1

(a) Differentiation and maturation scheme for early and late stage human ESC-CMs (RuES2) and iPSC-CM (IMR90). hESCs were maintained in a mouse embryonic fibroblast-conditioned medium (MEF-CM) supplemented with fibroblast growth factor (FGF). iPSCs were maintained in mTeSR. Both RuES2 and IMR90 stem cells were differentiated into CMs (day 0) by serial treatment with activin A and BMP-4. On day 14, hESC-CMs were transduced with MCK7-Cre lentivirus during replating onto PEI-gelatin coated glass coverslips/plastic plates. On day 19 cells were replated a second time in the presence of 2 ug/mL puromycin and selected for 48 hours. Cells were maintained for approximately 3 months in culture. We classified cells between days 25 and 40 as early stage PSC-CMs and cells > day 100 as late stage PSC-CMs. Alpha-actinin immunofluorescence (b and d) and mitochondrial staining (red)/eGFP (green) (c and e) of early stage (b-c) and late stage (d-e) hESC-CMs. (f–i) hESC-CM morphometric analysis. relative to early stage hESC-CMs, –121 cells.

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