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©The Author(s) 2021.
World J Stem Cells. Aug 26, 2021; 13(8): 1094-1111
Published online Aug 26, 2021. doi: 10.4252/wjsc.v13.i8.1094
Published online Aug 26, 2021. doi: 10.4252/wjsc.v13.i8.1094
Table 1 Protocols to produce induced pluripotent stem cell-derived mesenchymal stem cells by the mesenchymal stem cell Switch methods
Ref. | iPSC origin | iPSC to iMSC protocol | Time | Application | Citations |
Lian et al[41], 2010 | Lung fibroblast | (1) iPSC cultured on a gelatinized dish + KO DMEM + 10% SRM + bFGF + PDGFAB + EGF; and (2) FACS SORTING: CD24- and CD105+ and single cell clones plating | 7 | Limb ischemia in mice | 419 |
Giuliani et al[42], 2011 | Amniocytes | iPSC cultured 4 wk in DMEM/F12 + 10% hiFBS + b-FGF + NEAA + L-Glutamine + β-ME + p/s | 28 | Immunomodulatory properties of iMSC on NK cytolytic activity | 110 |
Liu et al[43], 2012 | Dermal fibroblast | (1) iPSC cultured on collagen-coated dishes with α-MEM + 10% FBS + dexamethasone + magnesium L-ascorbic acid phosphate + p/s; and (2) Cells cultured on collagen-coated dishes with α-MEM + 10% FBS + L-Glutamine + NEAA + p/s | 10 | iMSC generation w/ Fibrillar Collagen Coating | 118 |
Zou et al[44], 2013 | Dermal fibroblasts | iPSC medium switched for MSC medium: DMEM-low glucose + 10% FBS + L-Glutamine | 14 | Generation of osteogenesis 3D scaffolds | 98 |
Hynes et al[45], 2014 | Gingival fibroblast periodontal ligaments | (1) iPSC cultured with MSC medium: α-MEM + FCS + sodium pyruvate + l-ascorbate-2-phosphate + L-Glutamine + NEAA + HEPES + p/s; and (2) Cells cultured on gelatin-coated-flasks then switch to non-coated flasks | 14 | Generation of iMSC | 102 |
Jeong et al[46], 20141 | NA | (1) iPSCs cultured in iMSC-inducing medium: DMEM/F12 + 20% KOSR + SB431542 (TGFβ inhibitor); (2) EB grown on matrigel + DMEM/F12 + 0.5% BSA + 10% ITS + SB431542 (TGFβ inhibitor); and (3) Outgrowth grown with DMEM/F12 + 10% FBS + p/s | 17 | Duchene muscular dystrophy | 17 |
Hu et al[47], 2015 | iPS-S-01, C1P33, PCKDSF001C1 | iPSC medium switched for MSC medium: DMEM-low glucose + 10% FBS + L-Glutamine, then cultured in gelatin-coated dishes | 14 | Limb ischemia | 177 |
Kang et al[48], 2015 | Dermal fibroblasts | iPSC cultured with MSC medium: DMEM low glucose + FBS 10% + L-Glutamine + p/s then cultured on gelatin-coated dishes | 14 | iMSC plasticity (less adipogenesis) | 65 |
Zhang et al[23], 2015 | PBMCs | (1) iPSC medium switched for MSC medium: DMEM-low glucose + 10% FBS + L-Glutamine + NEAA + p/s; and (2) Cells cultured on gelatin-coated dishes | 17 | Cutaneous wound healing | 262 |
Lian et al[49], 2016 | NA | (1) iPSC cultured on gelatin-coated plates with MSC differentiation medium: KO DMEM + KOSR + bFGF + PDGFAB + EGF; and (2) FACS: CD24- CD105+ cells cultured on gelatin-coated plates with DMEM + 10% FBS + bFGF + PDGFAB + EGF | 20 | Directed differentiation of iPSC to MSC | 15 |
Gao et al[50], 2017 | Urine cell; Amniocytes | (1) iPSC cultured with MSC-inducing culture media: α-MEM + SRM + sodium pyruvate + l-ascorbate-2-phosphate + L-Glutamine + NEAA + p/s on gelatin-coated plates; and (2) Cells cultured with MSC maintenance medium = high-glucose DMEM + 10% FBS + bFGF + EGF | 17 | iMSC effect on dendritic cells | 27 |
Nachlas et al[51], 20181 | NA | (1) iPSC cultured in suspension (to promote cell aggregate) with differentiation media: KO-DMEM + β-ME+ L-Glutamine + 20% FBS + NEAA + p/s, then, cells were cultured on gelatin coated plates; and (2) Cells cultured with iMSC media: KO-DMEM + L-Glutamine + 10% FBS + NEAA + p/s | 12 | Generation of valve interstitial-like cells from iMSC | 14 |
Wang et al[52], 2018 | Amniocytes | (1) iPSC cultured with induction medium: α-MEM + 10% FBS + p/s + L-Glutamine + NEAA + sodium pyruvate + l-ascorbate-2- phosphate; (2) Cells plated on gelatin-coated plates; and (3) Cells plated on uncoated plates with iPSC-MSC medium = High-Glucose DMEM + FBS + bFGF + EGF + p/s | 14 | Effect of Dexamethasone on iMSC | 3 |
Wang et al[53], 2018 | PBMCs | iPSC cultured with MSC medium: Low-Glucose DMEM + 10% FBS + p/s + L-glutamine | NA | Immunomodulatory properties of MSC, transcriptome analysis | 8 |
McGrath et al[54], 20191 | Dermal fibroblast | (1) iPSC-MP thawed and expanded in KO DMEM + bFGF + L-Glutamine + MEM + NEAA + 20% FBS + Antibiotic-Antimycotic + β-ME; and (2) Cell are plated into gelatin coated-plates with KO DMEM + heparin + hPL + bFGF + L-Glutamine + MEM NEAA + Antibiotic-Antimycotic + β-ME | NA | iMSC differentiation: GMP-compatible and xeno-free cultivation | 6 |
Table 2 Protocols to produce induced pluripotent stem cell-derived mesenchymal stem cells by embryoid bodies approaches
Ref. | iPSC origin | iPSC to iMSC protocol | Time | Application | Citations |
Ahfeldt et al[56], 2012 | Foreskin fibroblast | (1) iPSC cultured into low-adhesion dishes for EB formation with DMEM + 15% FBS + L-Glutamine; (2) EB plated into gelatin-coated dishes with DMEM + 15% FBS + L-Glutamine; and (3) Cells plated with Mensenchymal Progenitor Cell (MCP) medium: DMEM + 15% FBS + L-Glutamine + bFGF | 12 | Producing white and brown adipocytes from hPSCs | 194 |
Chen et al[57], 20121 | Lung fibroblast | SB431542 inhibitor differentiation method (feeder free); iPSC cultured in inhibitor differentiation medium: KOSR medium + SB431542 (TGFβ inhibitor)Without bFGF to enhance differentiation. Embryoid body differentiation method: (1) EB formation in KOSR medium; and (2) EB cultured with MSC medium: DMEM + 10% FCS + L-Glutamine + gentamicin + p/s | 17 | Generation of iMSC with TGF-beta inhibitor | 136 |
Villa-Diaz et al[58], 2012 | Dermal fibroblasts | (1) EB formation in suspension cultured into ultra-low-attachment plates; and (2) EB plated on gelatin-coated dishes with MSC medium: α-MEM + 10% FBS + L-Glutamine + NEAA + FGF2 | 21 | iMSC from iPSC cultured on synthetic substrate (PMEDSAH) | 262 |
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 |
Shao et al[60], 2013 | MSC | (1) iPSC cultivated in suspension in the differentiation medium: KO DMEM + 20% FBS+ 1% NEAA + β-ME+ L-Glutamine for EB formation; and (2) Embryoid bodies plated on gelatin-coated dishes | 19 | iMSC DNA methylation profiles | 48 |
Jeong et al[46], 20141 | NA | (1) iPSCs cultured in iMSC-inducing medium: DMEM/F12 + 20% KOSR + SB431542 (TGFβ inhibitor); (2) EB grown on matrigel + DMEM/F12 + 0.5% BSA + 10% ITS + SB431542 (TGFβ inhibitor); and (3) Outgrowth grown with DMEM/F12 + FBS (10%) + p/s | 17 | Duchene muscular dystrophy | 17 |
Miao et al[61], 2014 | Dermal fibroblasts | EB cultured with DMEM + 10% FBS | NA | Myocardial infarctus | 38 |
Tang et al[21], 2014 | bone marrow | (1) iPSC cultured in ultra-low attachment plate to form EB with differentiation medium: DMEM/F12 + 20% KSR + MEF medium + NEAA + L-Glutamine + β-ME; and (2) EB plated into gelatin-coated plates + MSC growth medium: DMEM + 10% FBS + L-Glutamine + p/s | 20 | iMSC and calcium phosphate scaffold for bone regeneration | 88 |
Sheyn et al[20], 2016 | Dermal fibroblasts | (1) iPSC plated into PCR plates to form EB with IMDM medium: MDM media + KOSR + NEAA + β-ME + PSA antifungal-antibacterial solution; (2) EB transferred to poly-HEMA-coated flasks; (3) Attached EB (aiMSCs) and Transferred EB (tiMSCs) cultured into gelatin-coated flask with medium + TGF-β1; and (4) Medium switched for DMEM + 10% FBS + L-Glutamine + p/s | 10 | Generation of iMSC and repair bone defect | 60 |
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; and (3) FACS: PDGFR-α+ and VEGFR2+ cells resuspended on collagen-coated plates with αMEM + 10% FBS + 20% KOSR | 10 | iMSC from mesoderm or neuroepithelium differentiation | 7 |
Nachlas et al[51], 20181 | NA | (1) iPSC cultured in suspension (to promote cell aggregate) with differentiation media: KO-DMEM + β-ME + L-Glutamine + 20% FBS + NEAA + p/s, then cells were cultured on gelatin coated plates; and (2) Cells cultured with iMSC media: KO-DMEM + L-Glutamine + 10% FBS + NEAA + p/s | 12 | Generation of valve interstitial-like cells from iMSC | 14 |
Karam et al[62], 2020 | PBMC | (1) EB cultured into ultra-low attachment plates with differentiation medium: Low-Glucose DMEM + 15% FBS + p/s; (2) Later, RA is added to enhance EB formation; (3) EB plating into gelatin/matrigel coated plates + differentiation medium; and (4) Later bFGF is added | 14 | Generation of iMSC and adipocytes | NA |
Huang et al[63], 2020 | PBMC | (1) iPSC cultured in suspension to form EB; and (2) Cells plated into gelatin-coated plates with α-MEM + FGF2 | NA | Repair of acute kidney injury | NA |
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 |
Table 4 Protocols to produce induced pluripotent stem cell-derived mesenchymal stem cells by Pathway Inhibitor approaches
Ref. | iPSC origin | iPSC to iMSC protocol | Time | Application | Citations |
Chen YS et al[57], 20121 | Lung fibroblast | SB431542 Inhibitor Differentiation Method (feeder free); iPSC cultured in inhibitor differentiation medium: KOSR medium + SB431542 (TGFβ inhibitor)Without bFGF to enhance differentiation; Embryoid body differentiation method: (1) EB formation in KOSR medium; and (2) EB cultured with MSC medium: DMEM + 10% FCS + L-Glutamine + gentamicin + p/s | 17 | Generation of iMSC with TGF-beta inhibitor | 136 |
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 |
Jeong et al[46], 20141 | NA | (1) iPSCs cultured in iMSC-inducing medium: DMEM/F12 + 20% KOSR + SB431542 (TGFβ inhibitor); and (2) EB grown on matrigel + DMEM/F12 + 0.5% BSA + 10% ITS + SB431542. 3. Outgrowth grown with DMEM/F12 + 10% FBS + p/s | 17 | Duchene muscular dystrophy | 17 |
Zhao et al[67], 2015 | Blood cells | (1) iPSC cultured with mTeSR1 + SB431542 (TGFβ inhibitor) on matrigel-coated plates (7.5% CO2 atmosphere); and (2) Cells cultured with ESC–MSC medium: KO DMEM + KOSR + NEAA + p/s + L-Glutamine + β-ME + bFGF + EGF + SB431542 | 45 | Tumor tropism of iMSC | 79 |
Table 5 Protocols to produce induced pluripotent stem cell-derived mesenchymal stem cells by approaches that use Platelet Lysate
Ref. | iPSC origin | iPSC to iMSC protocol | Time | Application | Citations |
Frobel et al[71], 2014 | BM-MSCs | (1) EB formation on ultra-low attachment plates; and (2) Cells cultured with standard medium for MSC: DMEM + L-Glutamine + p/s + hPL + heparin on matrigel-coated wells then passaged on gelatin-coated wells | 35 | Epigenetic study of iMSC | 116 |
Luzzani et al[72], 2015 | Foreskin fibroblasts | (1) iPSC cultured in matrigel/geltrex-coated dishes with a-MEM + 10% PL + p/s + B7 or DMEM + 10% FBS; and (2) Cells cultured with no-coated dishes | 20 | MSC differentiation using platelet lysate | 26 |
McGrath et al[54], 20191 | Dermal fibroblasts | (1) iPSC-MP thawed and expanded in KO DMEM + bFGF + L-Glutamine + MEM NEAA + FBS (20%) + Antibiotic-Antimycotic+ β-ME; and (2) Cell are plated on gelatin coated-plates with DMEM KO + heparin + hPL+ bFGF + L-Glutamine + MEM NEAA + Antibiotic-Antimycotic+ β-ME | NA | iMSC differentiation: GMP-compatible and xeno-free cultivation | 6 |
Table 6 Frequency of use of iMSC surface markers
Positive CSM | % | Negative CSM | % |
CD73 | 18.1 | CD45 | 24.5 |
CD105 | 17.1 | CD34 | 23.0 |
CD90 | 15.7 | CD14 | 8.6 |
CD44 | 12.5 | CD31 | 7.2 |
CD29 | 9.3 | HLA-DR | 5.8 |
CD166 | 6.9 | CD11b | 5.0 |
CD146 | 3.7 | CD133 | 2.9 |
CD49(a) | 2.8 | TRA181 | 2.9 |
HLA-ABC | 2.3 | CD19 | 2.2 |
CD49(e) | 1.9 | CD24 | 2.2 |
CD106 | 1.4 | CD3 | 1.4 |
CD271 | 1.4 | CD40 | 1.4 |
CD49(d) | 0.9 | CD56 | 1.4 |
CD140alpha | 0.9 | CD80 | 1.4 |
Sca1 | 0.9 | CD86 | 1.4 |
CD33 | 0.5 | Oct3/4 | 1.4 |
CD49(f) | 0.5 | CD4 | 0.7 |
CD54 | 0.5 | CD20 | 0.7 |
CD71 | 0.5 | CD79a | 0.7 |
CD140(b) | 0.5 | CD117 | 0.7 |
CD144 | 0.5 | CD309 | 0.7 |
CD172alpha | 0.5 | Sox2 | 0.7 |
αSMA+ | 0.5 | TRA-160 | 0.7 |
Stro1 | 0.5 | TRA-161 | 0.7 |
TRA180 | 0.7 | ||
SSEA-4 | 0.7 |
- Citation: Dupuis V, Oltra E. Methods to produce induced pluripotent stem cell-derived mesenchymal stem cells: Mesenchymal stem cells from induced pluripotent stem cells. World J Stem Cells 2021; 13(8): 1094-1111
- URL: https://www.wjgnet.com/1948-0210/full/v13/i8/1094.htm
- DOI: https://dx.doi.org/10.4252/wjsc.v13.i8.1094