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Table 1 In vitro differentiation of human induced pluripotent stem cells (iPSCs) into red blood cells (RBCs)

From: Differentiation of human induced pluripotent stem cells into erythroid cells

Human iPSC cell source Reprogramming transcription factors Cluster of differentiation (CD) markers iPSC culture condition Results Refs.
IMR90 POU5F1, SOX2, and NANOG CD34+ and CD43+ (hematopoietic progenitors), CD31+ and CD43− (endothelial cells), CD43+, CD235a+, and CD41a+/− (erythro-megakaryopoietic) α-MEM with 20% defined FBS, 100 ng/mL bFGF, OP9 feeder layer Seven human iPSC lines could differentiate into RBCs with the similar pattern of differentiation [81]
Fetal and newborn foreskin fibroblasts POU5F1, SOX2, NANOG, and LIN28
Adult skin fibroblasts POU5F1, SOX2, and NANOG (M3-6) or POU5F1, SOX2, NANOG, and LIN28
IMR90 and FD-136 pSin-EF2-Oct4-Pur, pSin-EF2-Sox2-Pur, pSin-EF2-Nanog-Pur and pSin-EF2-Lin28-Pur13 CD34 and/or CD45 (hematopoietic progenitors), CD36 and CD235a (erythroid cells), CD71 (transferrin receptor), CD45, CD34, and CD71 (hematopoietic and erythroid cells) EB formation on a cellular stroma
100 ng/mL SCF, 100 ng/mL TPO, 100 ng/mL FL, 10 ng/mL BMP4, 5 ng/mL VEGF, 5 ng/mL IL-3, 5 ng/mL IL-6, 3 U/mL Epo, 10 μg/mL insulin, 3 U/mL heparin
The complete differentiation of human iPSCs into definitive erythrocytes and RBCs with fetal hemoglobin [33]
Human adult and fetal fibroblasts POU5F1, SOX2, and NANOG CD235a+ and CD45− (leukocyte-free RBCs), CD34+ or CD31+ (erythroid cells) 100 ng/mL ZbFGF, OP9 feeder layer, serum free medium, SCF, G-CSF, GM-CSF, IL3, IL6 The episomal reprogramming or transgene-free human iPSCs for large-scale expansion of RBCs [82]
Neonatal fibroblasts Episomal vectors that express OCT4, SOX2, NANOG, LIN28, MYC, KLF4, and LT
Human cord blood OCT4 and SOX2 alone (CD34-2F-iPSC) or expressing OCT4, SOX2, KLF4, and c-MYC (CD34-4F-iPSC) CD34+ (iPSCs), CD45+/CD34+ (HSCs), CD45+/CD34− (myeloid precursors), GPA+/CD45− (erythroid cells), CD36 and CD71 (primitive erythroid cells) 10% human plasma, 10 μg/mL insulin, 330 μg/mL human holotransferrin, 100 ng/mL SCF, 100 ng/mL TPO, 100 ng/mL Flt3-L, 5 ng/mL IL-3, 5 ng/mL IL-6, 5 ng/mL VEGF, 10–20 ng/mL BMP4, 3 U/mL EPO The growth rate of erythroid cells from iPSC-derived CD34+ HSCs was slightly higher [34]
iPSC line (33D6), iPSC lines from fibroblast cells (blood group O RhD2), and peripheral blood   CD144+/CD31+ (endothelial cells), CD31, CD34, CD36, CD41a, CD43, CD44, CD45, CD71, and CD235a Stemline II medium, 20 ng/mL bFGF, 20 ng/mL recombinant vitronectin, 1 mM StemRegenin (SR1), 1 mM hydrocortisone, 30–50 ng/mL SCF, 16.7 ng/mL Flt3-ligand, 10 ng/mL Wnt3A, 2 mM GSK3b inhibitor VIII or A-A014418, 6.7–20 ng/mL BMP4, 6.7 ng/mL IL-3, 6.7 ng/mL IL-11, 50 mM IBMX, 1.3 U/mL EPO, 30 ng/mL VEGF, 10 ng/mL FGFa, 10 ng/mL IGF, 10 ng/mL TPO, 5 mg/mL heparin, 50 mM IBMX, 0.4 ng/mL b-estradiol The large-scale expansion of human iPSC-derived erythroid cells under feeder-free and serum-free culture condition [83]
Cord blood CD34+ cells OCT4, SOX2, KLF4, and c-Myc CD43+ (hematopoietic progenitors), CD36, CD235a, CD45, CD71 (hematopoietic markers), CD31, CD144, CD41a, CD309, and CD4 VEGF, BMP4, Flt3-ligand, IL-3, IL-6, SCF, TPO, EPO Human iPSC-derived CD43-expressing hematopoietic cells are a suitable option for in vitro erythropoiesis [79]
PBMCs or MSCs from SCD patients Oct4, Klf4, Sox2, and c-Myc CD36+/CD71+ (peripheral blood erythroid progenitors (EP)), CD31, CD34, CD41a, CD43, CD45, CD71, CD73, CD144, CD235a, CD309 IDMEM medium, 0.2 mg/mL insulin, 0.11 mg/mL transferrin, 0.1 μg/mL sodium selenite, 0.45 mM a-mono-thioglycerol, 50 μg/mL AA, 20 ng/mL VEGF, 50 ng/mL SCF, 50 ng/ml fms-related tyrosine kinase 3 ligand, 50 ng/mL TPO, 5 μg/mL IL-3, 10 ng/mL BMP4,
5 U/ml EPO
MSC-derived iPSCs produced more efficient definitive erythroid cells with higher b-globin expression [48]
Human urine OCT4, SOX2, KLF4, and MYC CD34, CD43, CD45, CD31, CD144, CD235a, CD11b, CD14, CD3, CD4, CD5, CD7, CD8a Matrigel, mTeSR1 medium, stemline II, ITS, 20 ng/mL BMP4, 5 ng/mL Activin A, 5 ng/mL bFGF, 40 ng/mL VEGF, 50 ng/mL SCF, 50 ng/mL Flt3-ligand, 10 ng/mL TPO, 50 ng/mL IL-3, 50 ng/mL IL-6, UM171 improved in vitro derivation of HSCs from human iPSCs [84]
Cord blood CD34+ cells and CD36+ erythroblasts OCT4, SOX2, KLF4, and c- MYC CD34+/CD45+ (hematopoietic progenitors), CD36+/CD45+ (erythroid precursors) Matrigel, STEMdiff™ APEL™2 medium, 5% PFHM-II Protein-free Hybridoma Medium, 5 ng/mL IL-3, 100 ng/mL SCF, 3 U/mL EPO, 10% human plasma, 10 μg/mL insulin, 330 μg/mL human holotransferrin Prolonged human iPSC-derived RBCs in a simplified cell culture system with low cytokine support [31]
WT-iPSC line   CD34, CD38, CD45, CD90, CD117, CD133 Vitronectin, OP9 feeder layer, MEM medium with 10% FBS, 100 μM MTG, 50 μg/mL AA NOX4 has a significant role in the early stages of hematopoietic differentiation from iPSCs [85]
Bone marrow stromal cells from a SCD patient   CD31, CD34, CD36, CD38, CD41a, CD43, CD45, CD45RA, CD49f, CD71, CD73, CD90, CD144, CD184 mTeSR1 media, Matrigel, IMDM, C3H10T1/2 feeder cells, 1% ITS, 50 mg/mL AA, 0.45 mM a-monothioglycerol, 20 ng/mL human VEGF, 15% FBS or 20% KSR, OP9 feeder cells, 50 ng/mL FL, 50 ng/mL TPO, 5 ng/mL IL3, 50 ng/mL SCF, 5 U/mL EPO, and 10 ng/mL BMP4, 1.0 μM estradiol, 1.0 μM dexamethasone, 2% BSA, 0.56 mg/mL transferrin Serum-free iPSC sac-derived erythroid differentiation [38]
  1. IMR90 human fetal lung fibroblasts, PBMCs peripheral blood mononuclear cells, IDMEM Iscove’s modified Dulbecco’s medium, SCD sickle cell disease, FD-136 skin primary fibroblast cell line, OP9 mouse bone marrow stromal cell line, EB embryoid body, SCF stem cell factor, TPO thrombopoietin, FLT3 Fms-related tyrosine kinase 3 ligand, FL FLT3 ligand, BMP4 bone morphogenetic protein 4, VEGF vascular endothelial growth factor, IL-3 interleukin-3, EPO erythropoietin, ZbFGF zebrafish basic fibroblast growth factor, HSCs hematopoietic stem cells, IGF insulin-like growth factor, IBMX isobutyl methyl xanthine, MTG monothioglycerol, AA ascorbic acid, KSR knockout serum replacement, BSA bovine serum albumin, ITS insulin, transferrin, selenium