Methods to produce induced pluripotent stem cell-derived mesenchymal stem cells: Mesenchymal stem cells from induced pluripotent stem cells

Table 3 Protocols to produce induced pluripotent stem cell-derived mesenchymal stem cells by Specific Differentiation approaches

Ref.	iPSC origin	iPSC to iMSC protocol	Time	Application	Citations
Wei et al[59], 20121	Dermal fibroblasts	(1) EB formation through cardiac differentiation protocol involving cardiomyogenic medium CARM: High-Glucose DMEM + L-Glutamine + NEAA + Selenium Transferrin + β-ME + SB 203580 (p38-MAPK inhibitor); and (2) EB plating on gelatin-coated plates with DMEM + 2% FBS	21	Generation of iMSC	64
Fukuta et al[66], 20141	Dermal fibroblast	(1) Induction of hNCC from iPSC; (2) Cells cultured on fibronectin-coated dishes with STK2 medium + CDM (IMDM/Ham's F-12 + lipid concentrate + apo-transferrin + monothioglycerol + BSA + insulin + p/s) + SB431542 (TGFβ inhibitor) + CHIR (Wnt Agonist); and (3) Cells cultured with αMEM + 10% FBS	15	iMSC differentiation through neural crest lineage	80
Ouchi et al[65], 2016	Dermal fibroblast	(1) Generation of NCL (neural crest like-cells); and (2) NCL cultured into DMEM/F12 + Neurobasal medium + L-Glutamine + Gem21 Neuroplex + N2 Supplement + hbFGF + hEGF + insulin + p/s	10	iNCC can develop into iMSC	8
Eto et al[55], 20181	skin fibroblast	(1) iPSCs treated with CTK (collagenase type IV + 0.25% trypsin + KSR) and transferred to petri dishes to form EB with: DMEM/F12 + 20% KOSR + glutamine + NEAA + BMP4 + p/s; (2) Specific Differentiation or Mesodermal Differentiation: EB cultured on collagen-coated plates + αMEM + 10% FBS + bFGF + BMP4 + Activin A + LiCl + p/s or Neuroepithelial differentiation: αMEM + 10% FBS + β-ME + RA; (3) FACS: PDGFR-α+ and VEGFR2+ cells resuspended on collagen-coated plates with αMEM + 10% FBS + 20% KOSR	NA	iMSC from mesoderm or neuroepithelium differentiation	7
Mitsuzawa et al[64], 2019	NA	(1) Induction of hNCC: CDM (IMDM/Ham's F-12 + lipid concentrate + apo-transferrin+ monothioglycerol + BSA + insulin + p/s) + SB431542 (TGFβ inhibitor) + CHIR (Wnt Agonist)Maintenance in DMEM + 10% FBS + FGF2; and (2) Induction of iMSC with DMEM + 10% FBS + FGF2 on fibronectin coated plates	25	Hind limb in rat allotransplantation	1

iPSC origin refers to the cell type used for reprograming; Time is indicated as the minimum number of days required to obtain iMSCs; Citations show numbers on March 2020.

¹In references indicate the study includes methods in more than one protocol category. bFGF: Basic fibroblast growth factor; β-ME: β-Mercaptoethanol; BMP4: Bone morphogenetic protein 4; BSA: Bovine serum albumin; CARM: Cardiomyogenic medium; CDM: Chemical defined medium; CHIR: Wnt agonist, potent GSK3 inhibitor; DMEM/F12: Dulbecco's modified Eagle's medium EB: Embryoid body; FBS: Fetal bovine serum; FGF2: Fibroblast growth factor 2; Ham's F12: Medium formulated for single-cell plating of near-diploid Chinese hamster ovary cells; hbFGF: Human basic fibroblast growth factor; hEGF: Human epidermal growth factor; IMDM: Iscove's modified Dulbecco's media; iMSCs: Induced pluripotent stem cell-derived mesenchymal stem cells; iPSC: Induced pluripotent stem cell; ITS: Insulin-transferrin-selenium; KOSR: Knock-out serum replacement; KSR: Knockout serum replacement; NA: Not available; NEAA: Non-essential amino acid; α-MEM: Minimum essential medium Eagle; PDGFR: Platelet-derived growth factor receptor A; p/s: Penicillin-streptomycin; RA: Retinoic acid; TGF: Transforming growth factor; VEGFR: Vascular endothelial growth factor.