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Generation of Hematopoietic Stem and Progenitor Cells from Human Pluripotent Stem Cells

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In Vitro Differentiation of T-Cells

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2048))

Abstract

Human pluripotent stem cells (PSCs) have the potential to provide a virtually unlimited supply of cells for transplantation therapy. When combined with recent advances in genome editing technologies, human PSCs could offer various approaches that enable gene therapy, drug discovery, disease modeling, and in vitro modeling of human development. De novo generation of hematopoietic stem cells (HSCs) from human PSCs is an important focus in the field, since it enables autologous HSC transplantation to treat many blood disorders and malignancies. Although culture conditions have been established to generate a broad spectrum of hematopoietic progenitors from human PSCs, it remains a significant challenge to generate bona fide HSCs that possess sustained self-renewal and multilineage differentiation capacities upon transplantation. In this review, recent promising advances in the efforts to generate HSCs and hematopoietic progenitors from human PSCs in vitro and in vivo or from somatic cells are discussed.

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References

  1. Orkin SH, Zon LI (2008) Hematopoiesis: an evolving paradigm for stem cell biology. Cell 132(4):631–644

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  2. Niederwieser D, Baldomero H, Szer J, Gratwohl M, Aljurf M, Atsuta Y et al (2016) Hematopoietic stem cell transplantation activity worldwide in 2012 and a SWOT analysis of the Worldwide Network for Blood and Marrow Transplantation Group including the global survey. Bone Marrow Transplant 51(6):778–785

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  3. Giralt S, Costa L, Schriber J, Dipersio J, Maziarz R, McCarty J et al (2014) Optimizing autologous stem cell mobilization strategies to improve patient outcomes: consensus guidelines and recommendations. Biol Blood Marrow Transplant 20(3):295–308

    Article  PubMed  Google Scholar 

  4. Kumar S, Geiger H (2017) HSC niche biology and hsc expansion ex vivo. Trends Mol Med 23(9):799–819

    Article  PubMed  PubMed Central  Google Scholar 

  5. Palis J, Robertson S, Kennedy M, Wall C, Keller G (1999) Development of erythroid and myeloid progenitors in the yolk sac and embryo proper of the mouse. Development 126(22):5073–5084

    CAS  PubMed  Google Scholar 

  6. Gomez Perdiguero E, Klapproth K, Schulz C, Busch K, Azzoni E, Crozet L et al (2015) Tissue-resident macrophages originate from yolk-sac-derived erythro-myeloid progenitors. Nature 518(7540):547–551

    Article  PubMed  CAS  Google Scholar 

  7. McGrath KE, Frame JM, Fegan KH, Bowen JR, Conway SJ, Catherman SC et al (2015) Distinct sources of hematopoietic progenitors emerge before HSCs and provide functional blood cells in the mammalian embryo. Cell Rep 11(12):1892–1904

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  8. Boiers C, Carrelha J, Lutteropp M, Luc S, Green JC, Azzoni E et al (2013) Lymphomyeloid contribution of an immune-restricted progenitor emerging prior to definitive hematopoietic stem cells. Cell Stem Cell 13(5):535–548

    Article  PubMed  CAS  Google Scholar 

  9. Medvinsky A, Dzierzak E (1996) Definitive hematopoiesis is autonomously initiated by the AGM region. Cell 86(6):897–906

    Article  CAS  PubMed  Google Scholar 

  10. de Bruijn MF, Ma X, Robin C, Ottersbach K, Sanchez MJ, Dzierzak E (2002) Hematopoietic stem cells localize to the endothelial cell layer in the midgestation mouse aorta. Immunity 16(5):673–683

    Article  PubMed  Google Scholar 

  11. de Bruijn MF, Speck NA, Peeters MC, Dzierzak E (2000) Definitive hematopoietic stem cells first develop within the major arterial regions of the mouse embryo. EMBO J 19(11):2465–2474

    Article  PubMed  PubMed Central  Google Scholar 

  12. Ottersbach K, Dzierzak E (2005) The murine placenta contains hematopoietic stem cells within the vascular labyrinth region. Dev Cell 8(3):377–387

    Article  CAS  PubMed  Google Scholar 

  13. Li Z, Lan Y, He W, Chen D, Wang J, Zhou F et al (2012) Mouse embryonic head as a site for hematopoietic stem cell development. Cell Stem Cell 11(5):663–675

    Article  CAS  PubMed  Google Scholar 

  14. Gekas C, Dieterlen-Lievre F, Orkin SH, Mikkola HK (2005) The placenta is a niche for hematopoietic stem cells. Dev Cell 8(3):365–375

    Article  CAS  PubMed  Google Scholar 

  15. Kaufman DS, Hanson ET, Lewis RL, Auerbach R, Thomson JA (2001) Hematopoietic colony-forming cells derived from human embryonic stem cells. Proc Natl Acad Sci U S A 98(19):10716–10721

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  16. Nakano T, Kodama H, Honjo T (1994) Generation of lymphohematopoietic cells from embryonic stem cells in culture. Science 265(5175):1098–1101

    Article  CAS  PubMed  Google Scholar 

  17. Vodyanik MA, Bork JA, Thomson JA, Slukvin II (2005) Human embryonic stem cell-derived CD34+ cells: efficient production in the coculture with OP9 stromal cells and analysis of lymphohematopoietic potential. Blood 105(2):617–626

    Article  CAS  PubMed  Google Scholar 

  18. Itskovitz-Eldor J, Schuldiner M, Karsenti D, Eden A, Yanuka O, Amit M et al (2000) Differentiation of human embryonic stem cells into embryoid bodies compromising the three embryonic germ layers. Mol Med 6(2):88–95

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  19. Chadwick K, Wang L, Li L, Menendez P, Murdoch B, Rouleau A et al (2003) Cytokines and BMP-4 promote hematopoietic differentiation of human embryonic stem cells. Blood 102(3):906–915

    Article  CAS  PubMed  Google Scholar 

  20. Cerdan C, Rouleau A, Bhatia M (2004) VEGF-A165 augments erythropoietic development from human embryonic stem cells. Blood 103(7):2504–2512

    Article  CAS  PubMed  Google Scholar 

  21. Pick M, Azzola L, Mossman A, Stanley EG, Elefanty AG (2007) Differentiation of human embryonic stem cells in serum-free medium reveals distinct roles for bone morphogenetic protein 4, vascular endothelial growth factor, stem cell factor, and fibroblast growth factor 2 in hematopoiesis. Stem Cells 25(9):2206–2214

    Article  CAS  PubMed  Google Scholar 

  22. Galic Z, Kitchen SG, Kacena A, Subramanian A, Burke B, Cortado R et al (2006) T lineage differentiation from human embryonic stem cells. Proc Natl Acad Sci U S A 103(31):11742–11747

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  23. Schmitt TM, Zuniga-Pflucker JC (2002) Induction of T cell development from hematopoietic progenitor cells by delta-like-1 in vitro. Immunity 17(6):749–756

    Article  CAS  PubMed  Google Scholar 

  24. Timmermans F, Velghe I, Vanwalleghem L, De Smedt M, Van Coppernolle S, Taghon T et al (2009) Generation of T cells from human embryonic stem cell-derived hematopoietic zones. J Immunol 182(11):6879–6888

    Article  CAS  PubMed  Google Scholar 

  25. Ramos-Mejia V, Melen GJ, Sanchez L, Gutierrez-Aranda I, Ligero G, Cortes JL et al (2010) Nodal/Activin signaling predicts human pluripotent stem cell lines prone to differentiate toward the hematopoietic lineage. Mol Ther 18(12):2173–2181

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  26. Inman GJ, Nicolas FJ, Callahan JF, Harling JD, Gaster LM, Reith AD et al (2002) SB-431542 is a potent and specific inhibitor of transforming growth factor-beta superfamily type I activin receptor-like kinase (ALK) receptors ALK4, ALK5, and ALK7. Mol Pharmacol 62(1):65–74

    Article  CAS  PubMed  Google Scholar 

  27. Kennedy M, Awong G, Sturgeon CM, Ditadi A, LaMotte-Mohs R, Zuniga-Pflucker JC et al (2012) T lymphocyte potential marks the emergence of definitive hematopoietic progenitors in human pluripotent stem cell differentiation cultures. Cell Rep 2(6):1722–1735

    Article  CAS  PubMed  Google Scholar 

  28. Woll PS, Morris JK, Painschab MS, Marcus RK, Kohn AD, Biechele TL et al (2008) Wnt signaling promotes hematoendothelial cell development from human embryonic stem cells. Blood 111(1):122–131

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  29. Wang Y, Nakayama N (2009) WNT and BMP signaling are both required for hematopoietic cell development from human ES cells. Stem Cell Res 3(2-3):113–125

    Article  PubMed  Google Scholar 

  30. Sturgeon CM, Ditadi A, Awong G, Kennedy M, Keller G (2014) Wnt signaling controls the specification of definitive and primitive hematopoiesis from human pluripotent stem cells. Nat Biotechnol 32(6):554–561

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  31. Choi KD, Vodyanik MA, Togarrati PP, Suknuntha K, Kumar A, Samarjeet F et al (2012) Identification of the hemogenic endothelial progenitor and its direct precursor in human pluripotent stem cell differentiation cultures. Cell Rep 2(3):553–567

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  32. Ditadi A, Sturgeon CM, Tober J, Awong G, Kennedy M, Yzaguirre AD et al (2015) Human definitive haemogenic endothelium and arterial vascular endothelium represent distinct lineages. Nat Cell Biol 17(5):580–591

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  33. Ackermann M, Liebhaber S, Klusmann JH, Lachmann N (2015) Lost in translation: pluripotent stem cell-derived hematopoiesis. EMBO Mol Med 7(11):1388–1402

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  34. Tian X, Woll PS, Morris JK, Linehan JL, Kaufman DS (2006) Hematopoietic engraftment of human embryonic stem cell-derived cells is regulated by recipient innate immunity. Stem Cells 24(5):1370–1380

    Article  CAS  PubMed  Google Scholar 

  35. Ledran MH, Krassowska A, Armstrong L, Dimmick I, Renstrom J, Lang R et al (2008) Efficient hematopoietic differentiation of human embryonic stem cells on stromal cells derived from hematopoietic niches. Cell Stem Cell 3(1):85–98

    Article  CAS  PubMed  Google Scholar 

  36. Shultz LD, Lyons BL, Burzenski LM, Gott B, Chen X, Chaleff S et al (2005) Human lymphoid and myeloid cell development in NOD/LtSz-scid IL2R gamma null mice engrafted with mobilized human hemopoietic stem cells. J Immunol 174(10):6477–6489

    Article  CAS  PubMed  Google Scholar 

  37. Lu M, Kardel MD, O'Connor MD, Eaves CJ (2009) Enhanced generation of hematopoietic cells from human hepatocarcinoma cell-stimulated human embryonic and induced pluripotent stem cells. Exp Hematol 37(8):924–936

    Article  CAS  PubMed  Google Scholar 

  38. Ito M, Hiramatsu H, Kobayashi K, Suzue K, Kawahata M, Hioki K et al (2002) NOD/SCID/gamma(c)(null) mouse: an excellent recipient mouse model for engraftment of human cells. Blood 100(9):3175–3182

    Article  CAS  PubMed  Google Scholar 

  39. Sauvageau G, Lansdorp PM, Eaves CJ, Hogge DE, Dragowska WH, Reid DS et al (1994) Differential expression of homeobox genes in functionally distinct CD34+ subpopulations of human bone marrow cells. Proc Natl Acad Sci U S A 91(25):12223–12227

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  40. Antonchuk J, Sauvageau G, Humphries RK (2002) HOXB4-induced expansion of adult hematopoietic stem cells ex vivo. Cell 109(1):39–45

    Article  CAS  PubMed  Google Scholar 

  41. Kyba M, Perlingeiro RC, Daley GQ (2002) HoxB4 confers definitive lymphoid-myeloid engraftment potential on embryonic stem cell and yolk sac hematopoietic progenitors. Cell 109(1):29–37

    Article  CAS  PubMed  Google Scholar 

  42. Wang L, Menendez P, Shojaei F, Li L, Mazurier F, Dick JE et al (2005) Generation of hematopoietic repopulating cells from human embryonic stem cells independent of ectopic HOXB4 expression. J Exp Med 201(10):1603–1614

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  43. Ramos-Mejia V, Navarro-Montero O, Ayllon V, Bueno C, Romero T, Real PJ et al (2014) HOXA9 promotes hematopoietic commitment of human embryonic stem cells. Blood 124(20):3065–3075

    Article  CAS  PubMed  Google Scholar 

  44. Dou DR, Calvanese V, Sierra MI, Nguyen AT, Minasian A, Saarikoski P et al (2016) Medial HOXA genes demarcate haematopoietic stem cell fate during human development. Nat Cell Biol 18(6):595–606

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  45. Marshall H, Morrison A, Studer M, Popperl H, Krumlauf R (1996) Retinoids and hox genes. FASEB J 10(9):969–978

    Article  CAS  PubMed  Google Scholar 

  46. Chanda B, Ditadi A, Iscove NN, Keller G (2013) Retinoic acid signaling is essential for embryonic hematopoietic stem cell development. Cell 155(1):215–227

    Article  CAS  PubMed  Google Scholar 

  47. Doulatov S, Vo LT, Chou SS, Kim PG, Arora N, Li H et al (2013) Induction of multipotential hematopoietic progenitors from human pluripotent stem cells via respecification of lineage-restricted precursors. Cell Stem Cell 13(4):459–470

    Article  CAS  PubMed  Google Scholar 

  48. Sugimura R, Jha DK, Han A, Soria-Valles C, da Rocha EL, Lu YF et al (2017) Haematopoietic stem and progenitor cells from human pluripotent stem cells. Nature 545(7655):432–438

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  49. Tan YT, Ye L, Xie F, Beyer AI, Muench MO, Wang J et al (2018) Respecifying human iPSC-derived blood cells into highly engraftable hematopoietic stem and progenitor cells with a single factor. Proc Natl Acad Sci U S A 115(9):2180–2185

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  50. Sanjuan-Pla A, Bueno C, Prieto C, Acha P, Stam RW, Marschalek R et al (2015) Revisiting the biology of infant t(4;11)/MLL-AF4+ B-cell acute lymphoblastic leukemia. Blood 126(25):2676–2685

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  51. Cudennec C, Nicolas JF (1977) Blood formation in a clonal cell line of mouse teratocarcinoma. J Embryol Exp Morphol 38:203–210

    CAS  PubMed  Google Scholar 

  52. Cudennec CA, Johnson GR (1981) Presence of multipotential hemopoietic cells in teratocarcinoma cultures. J Embryol Exp Morphol 61:51–59

    CAS  PubMed  Google Scholar 

  53. Cudennec CA, Salaun J (1979) Definitive red blood cell differentiation in a clonal line of mouse teratocarcinoma cultured in vivo in the chick embryo. Cell Differ 8(2):75–82

    Article  CAS  PubMed  Google Scholar 

  54. Amabile G, Welner RS, Nombela-Arrieta C, D'Alise AM, Di Ruscio A, Ebralidze AK et al (2013) In vivo generation of transplantable human hematopoietic cells from induced pluripotent stem cells. Blood 121(8):1255–1264

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  55. Suzuki N, Yamazaki S, Yamaguchi T, Okabe M, Masaki H, Takaki S et al (2013) Generation of engraftable hematopoietic stem cells from induced pluripotent stem cells by way of teratoma formation. Mol Ther 21(7):1424–1431

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  56. Tsukada M, Ota Y, Wilkinson AC, Becker HJ, Osato M, Nakauchi H et al (2017) In vivo generation of engraftable murine hematopoietic stem cells by Gfi1b, c-Fos, and Gata2 overexpression within teratoma. Stem Cell Reports 9(4):1024–1033

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  57. Pereira CF, Chang B, Qiu J, Niu X, Papatsenko D, Hendry CE et al (2013) Induction of a hemogenic program in mouse fibroblasts. Cell Stem Cell 13(2):205–218

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  58. Batta K, Florkowska M, Kouskoff V, Lacaud G (2014) Direct reprogramming of murine fibroblasts to hematopoietic progenitor cells. Cell Rep 9(5):1871–1884

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  59. Cheng H, Ang HY, AEF C, Li P, Fang HT, Liu TM et al (2016) Reprogramming mouse fibroblasts into engraftable myeloerythroid and lymphoid progenitors. Nat Commun 7:13396

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  60. Sandler VM, Lis R, Liu Y, Kedem A, James D, Elemento O et al (2014) Reprogramming human endothelial cells to haematopoietic cells requires vascular induction. Nature 511(7509):312–318

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  61. Lis R, Karrasch CC, Poulos MG, Kunar B, Redmond D, Duran JGB et al (2017) Conversion of adult endothelium to immunocompetent haematopoietic stem cells. Nature 545(7655):439–445

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  62. Cosgun KN, Rahmig S, Mende N, Reinke S, Hauber I, Schafer C et al (2014) Kit regulates HSC engraftment across the human-mouse species barrier. Cell Stem Cell 15(2):227–238

    Article  CAS  PubMed  Google Scholar 

  63. McIntosh BE, Brown ME, Duffin BM, Maufort JP, Vereide DT, Slukvin II et al (2015) Nonirradiated NOD,B6.SCID Il2rgamma−/− Kit(W41/W41) (NBSGW) mice support multilineage engraftment of human hematopoietic cells. Stem Cell Reports. 4(2):171–180

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  64. Yurino A, Takenaka K, Yamauchi T, Nunomura T, Uehara Y, Jinnouchi F et al (2016) Enhanced reconstitution of human erythropoiesis and thrombopoiesis in an immunodeficient mouse model with kit(Wv) mutations. Stem Cell Reports. 7(3):425–438

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  65. Wunderlich M, Chou FS, Link KA, Mizukawa B, Perry RL, Carroll M et al (2010) AML xenograft efficiency is significantly improved in NOD/SCID-IL2RG mice constitutively expressing human SCF, GM-CSF and IL-3. Leukemia 24(10):1785–1788

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  66. Rongvaux A, Willinger T, Martinek J, Strowig T, Gearty SV, Teichmann LL et al (2014) Development and function of human innate immune cells in a humanized mouse model. Nat Biotechnol 32(4):364–372

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  67. Lan P, Tonomura N, Shimizu A, Wang S, Yang YG (2006) Reconstitution of a functional human immune system in immunodeficient mice through combined human fetal thymus/liver and CD34+ cell transplantation. Blood 108(2):487–492

    Article  CAS  PubMed  Google Scholar 

  68. Melkus MW, Estes JD, Padgett-Thomas A, Gatlin J, Denton PW, Othieno FA et al (2006) Humanized mice mount specific adaptive and innate immune responses to EBV and TSST-1. Nat Med 12(11):1316–1322

    Article  CAS  PubMed  Google Scholar 

  69. Sun X, Xu J, Lu H, Liu W, Miao Z, Sui X et al (2013) Directed differentiation of human embryonic stem cells into thymic epithelial progenitor-like cells reconstitutes the thymic microenvironment in vivo. Cell Stem Cell 13(2):230–236

    Article  CAS  PubMed  Google Scholar 

  70. Parent AV, Russ HA, Khan IS, LaFlam TN, Metzger TC, Anderson MS et al (2013) Generation of functional thymic epithelium from human embryonic stem cells that supports host T cell development. Cell Stem Cell 13(2):219–229

    Article  CAS  PubMed  Google Scholar 

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Acknowledgments

This work was supported by The Jackson Laboratory. The author thanks William Skarnes for thoughtful reading and editing of the manuscript and apologizes to authors whose papers could not be cited in this review owing to space limitations.

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  1. Cellular Engineering, The Jackson Laboratory for Genomic Medicine, Farmington, CT, USA

    Hideyuki Oguro

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  1. Hideyuki Oguro
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Correspondence to Hideyuki Oguro .

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  1. Shin Kaneko Laboratory, Department of Cell Growth and Differentiation, Center for iPS Cell Research and Application, Kyoto University, Kyoto, Japan

    Shin Kaneko

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Oguro, H. (2019). Generation of Hematopoietic Stem and Progenitor Cells from Human Pluripotent Stem Cells. In: Kaneko, S. (eds) In Vitro Differentiation of T-Cells. Methods in Molecular Biology, vol 2048. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-9728-2_19

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  • DOI: https://doi.org/10.1007/978-1-4939-9728-2_19

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  • Publisher Name: Humana, New York, NY

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  • Online ISBN: 978-1-4939-9728-2

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