Table 1.

human iPSC step-wise differentiation

Study	Induction factor	Medium	Duration of muscle commitment and differentiation (days)	Cell composition	Purification	In vitro	In vivo
Chal et al., 2015	days 0–6 CHIR99021 and LDN193189, insulin-transferrin-selenium; days 3–6 addition of FGF-2; days 6–8 IGF-1, HGF, FGF2 and LDN193189; days 8–12 IGF-1; days 12–50 IGF-1 and HGF	DMEM based medium; days 0–6 serum free, thereafter + knock-out serum replacement (KSR)	20–30	~22% of nuclei were MYOG+, and 23% of nuclei were PAX7+	No purification	MyHC+, MYOG+ fibers, PAX7+ cells, organized sarcomeres, with the periodic distribution of the sarcomeric proteins Titin and fast MyHC	No
Chal et al., 2016	days 0–6 CHIR99021 and LDN193189, insulin-transferrin-selenium; days 3–6 addition of FGF-2; days 6–8 IGF-1, HGF, FGF2 and LDN193189; days 8–12 IGF-1; days 12–50 IGF-1 and HGF	DMEM based medium; days 0–6 serum free, thereafter + KSR	30	Most MYOG+ cells, MyHC+ myofibers. PAX7+ satellite-like cells.	No purification	Sub-culturing of human myo-progenitors by day 28. MyHC+, MYOG+ fibers, PAX7+ cells, organized sarcomeres. Fast MyHC+ and h-Dystrophin+ fibers in muscle construct.	No
Choi et al., 2016	days 0–4 CHIR99021; days 4–12 DAPT	Serum-free N2 medium	30	~15% NCAM+ HNK1-myoblasts, 61.5% MYOG+ and 63.6% MyHC+ cells	NCAM+ HNK1-	MyHC, TITIN, DES, DYSTROPHIN, α-ACTININ	PAX7+ cells, h-Dystrophin+ fibers
Hicks et al., 2018	days 0–2 CHIR99021; days 12–20 FGF2, TGF-β inhibitor when in differentiation condition	days 0–12, days 20–35 E6 medium, days 12–20 StemPro-34 medium, days 35–50 DMEM/F12 + 1% ITS medium	30–50	PAX7, MYF5, MYOD, MYOGENIN and spontaneously contracting myotubes	ERBB3+NGFR+	Enriched for PAX7 and MYF5	h-LaminA/C+, h-Dystrophin+ fibers. In vivo engraftment of ERBB3+ hiPSC-SMPCs restored dystrophin to levels approaching uncultured fetal muscle
Sakai-Takemura et al., 2018	days 0–12 Chal et al. 2016 method; days 12–42 floating culture; days 42–70 adhesion culture	days 0–12 DMEM, days 12–70 10% FBS/DMEM	70	MYOGENIN+ myotubes	CD57(−) CD108(−) CD271(+) ERBB3(+) cells	MYOGENIN, MyHC	h-LaminA/C+ (nuclear membrane) and h-Spectrin+ (sarcolemma) myofibers
Wu et al., 2018	days 1–4 CHIR99021, BMP inhibitor, TGF-β inhibitor; days 5–15 BMP inhibitor, TGF-β inhibitor	MDM-I medium days1–4, MDM-II medium day 5–15, MDM-III medium for terminal differentiation	15	at day 4 99.4% PAX7+, at day 15, 50–55% MYF5+	CD10+CD24-	MyHC+ myotubes	myofibers expressing human markers (maximum of 50–60 fibers positive for h-Dystrophin and h-Lamin A/C)