Table 2:

Methods for the differentiation of hPSC into myogenic progenitors and in vivo engraftment

Method	Purification	In vivo experiment	Engraftment and function	Transgene	Reference
Adenoviral based delivery of MYOD1; monolayer cultures	N/A	Transplantation of 0.5 × 106 cells per TA muscle of immunodeficient Rag/mdx mice	150–175 fibers positive for human SPECTRIN per section	MYOD	[27]
Lentiviral delivery of MYOD1 in PSC-derived mesoangioblasts	N/A	Transplantation of 106 cells per TA muscle of immunodeficient Sgca-null/scid/beige mice	50 fibers positive for α-sarcoglycan per section	MYOD	[28]
Lentiviral delivery of doxycycline- inducible PAX7; embryoid body cultures	Purification of PAX7+ myogenic progenitors based on GFP transgene	Transplantation of 0.3 × 106 cells per TA muscle of immunodeficient NSG-mdx4Cv mice	100 fibers positive for human dystrophin per section. Also showed satellite cell engraftment and functional improvement of transplanted muscles in comparison with untreated muscles.	PAX7	[32]
Similar to [32]	Positive selection for surface markers CD54, integrin α9β1, and Syndecan2 for the purification of PAX7+ myogenic progenitors	Transplantation of 0.5 × 106 cells per in TA muscles of immunodeficient NSG and NSGmdx4Cv mice	50 fibers positive for human dystrophin per section. Also showed satellite cell engraftment	PAX7	[33]
hESC differentiated into mesenchymal precursor and then into myogenic progenitors	Positive selection for CD73 and NCAM	Transplantation of 0.5 × 106 cells per TA muscle of immunodeficient SCID/Beige mice	Few fibers positive for human nuclear antigen and human laminin positive fibers were detected	Transgene- free	[34]
hPSC differentiated into embryoid bodies and then into myogenic mesenchymal progenitor cells.	N/A	Transplantation of 0.5 × 106 cells per TA muscle of immunodeficient NOG mice	10–20% fibers containing human nuclei. Also showed satellite cell engraftment	Transgene- free	[35]
Monolayer culture based on the use of GSK3β inhibitor and FGF2 growth factor treatment	Positive selection for surface markers CXCR4 and CMET, and negative selection for ACHR and HNK1	N/A	N/A	Transgene- free	[36]
Similar to [36]	Positive selection for C-MET and negative selection for HNK1	Transplantation of 0.5 × 106 cells per TA muscle of immunodeficient NSG mice	50 fibers positive for human spectrin per section	Transgene-free	[38]
Similar to [36]	N/A	N/A	N/A	Transgene- free	[37]
Monolayer culture based on the use of GSK3β and BMP inhibition, and treatment with FGF2, HGF and IGF1	N/A	N/A	N/A	Transgene- free	[39, 40]
Embryoid body cultures and microRNA cocktail treatment	Positive selection for surface markers CD140a, CD140b, and CD44	Transplantation of 0.5 × 106 cells in the femoral artery of immunodeficient Rag2/Il2rg null Sgcb null mice	15–20% fibers positive for human dystrophin. Functional improvement shown in the Extensor Digitorum Longus (EDL) muscle	Transgene- free	[42, 43]
Similar to protocols in [37, 39]	Positive selection for surface markers ERBB3 and NGFR	Transplantation of 106 cells per TA muscles of immunodeficient mdx-NSG mice	150 fibers positive for human dystrophin/spectrin upon co-injection of cells with TGFβ inhibitor	Transgene- free	[44]
Monolayer culture based on the use of GSK3β and NOTCH inhibition	Positive selection for NCAM, and negative selection for HNK1	Transplantation of 1–3 × 106 cells per TA muscles of immunodeficient NOD-Rag1null IL2rgnull and NSG-mdx4cv mice	Approximately 100 fibers positive for human laminin and lamin A/C per section in NSGmdx4cv, and 200300 fibers in NOD-Rag1null IL2rgnull mice. Also showed satellite cell engraftment	Transgene- free	[45]
Monolayer culture based on the use of GSK3β, BMP and TGFβ inhibition	Positive selection for CD10, and negative selection for CD24	Transplantation of 0.25 × 106 cells per TA muscles of in immunodeficient NSG-mdx4cv mice	Approximately 50 fibers positive for human dystrophin/lamin A/C per section. Also showed satellite cell engraftment	Transgene- free	[46–48]