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] |