Table 1 Summary of the 15 differentiation conditions.

Condition	hPSC line	Seeding density (cells/well)	CHIR99021 concentration	Differentiation efficiency
1.5 m–12 μM	H9 ESC	1.5 × 10⁶	12 μM	0.3 ± 0.1%
1.5 m–9 μM	H9 ESC	1.5 × 10⁶	9 μM	0.5 ± 0.2%
100 k–12 μM	H9 ESC	1.0 × 10⁵	12 μM	0.6 ± 0.3%
19-9-11-600 k–8 μM	19-9-11 iPSC	6.0 × 10⁵	8 μM	5.9 ± 0.7%
H13-600 k–10 μM	H13 ESC	6.0 × 10⁵	8 μM	10.8 ± 1.3%
2 m–12 μM	H9 ESC	2.0 × 10⁶	12 μM	15.1 ± 1.3%
500 k–12 μM	H9 ESC	5.0 × 10⁵	12 μM	19.6 ± 1.5%
1 m–12 μM	H9 ESC	1.0 × 10⁶	12 μM	21.7 ± 2.3%
IMR90-1 m–12 μM	IMR90-4 iPSC	1.0 × 10⁶	12 μM	26.4 ± 1.2%
IMR90-1 m–9 μM	IMR90-4 iPSC	1.0 × 10⁶	9 μM	38.5 ± 1.9%
500 K–10 μM	H9 ESC	5.0 × 10⁵	10 μM	51.8 ± 3.2%
IMR90-1 m–10 μM	IMR90-4 iPSC	1.0 × 10⁶	10 μM	53.2 ± 0.8%
19-9-11-600 k–6 μM	19-9-11 iPSC	6.0 × 10⁵	6 μM	61.0 ± 3.2%
H13-800 k–10 μM	H13 ESC	8.0 × 10⁵	10 μM	63.1 ± 2.4%
500 K–9 μM	H9 ESC	5.0 × 10⁵	9 μM	65.5 ± 2.1%

hPSCs, including H9 and H13 embryonic stem cells (ESC) or IMR90-4 and 19-9-11 induced pluripotent stem cells (iPSC) were differentiated into CMs following an established method¹¹. H13 ESC, IMR904- iPSC, and 19-9-11 iPSC are specified with H13, IMR90, and 19-9-11. On differentiation day 12, cells were verified by flow cytometry with cTnT labeling from three independent replicates to define differentiation efficiency. Data were collected from three biological replicates. Conditions are presented with condition name (seeding density, CHIR99021 concentration, IMR90 status), hPSC line, seeding density, CHIR99021 (Wnt activator) concentration, and differentiation efficiency (mean ± SEM). Low differentiation efficiencies (< 50% cTnT+ on day 12) are in italic and high differentiation efficiencies (≥ 50% cTnT+ on day 12) are in bold.

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