The self-renewal capacity of multipotent haematopoietic stem cells (HSCs) supports blood system homeostasis throughout life and underlies the curative capacity of clinical HSC transplantation therapies. However, despite extensive characterization of the HSC state in the adult bone marrow and embryonic fetal liver, the mechanism of HSC self-renewal has remained elusive. This Review presents our current understanding of HSC self-renewal in vivo and ex vivo, and discusses important advances in ex vivo HSC expansion that are providing new biological insights and offering new therapeutic opportunities.
Subscribe to Journal
Get full journal access for 1 year
only $4.92 per issue
All prices are NET prices.
VAT will be added later in the checkout.
Tax calculation will be finalised during checkout.
Rent or Buy article
Get time limited or full article access on ReadCube.
All prices are NET prices.
Orkin, S. H. & Zon, L. I. Hematopoiesis: an evolving paradigm for stem cell biology. Cell 132, 631–644 (2008).
Gordon, M. Y., Lewis, J. L. & Marley, S. B. Of mice and men… and elephants. Blood 100, 4679–4680 (2002).
Eaves, C. J. Hematopoietic stem cells: concepts, definitions, and the new reality. Blood 125, 2605–2613 (2015).
Weissman, I. L. & Shizuru, J. A. The origins of the identification and isolation of hematopoietic stem cells, and their capability to induce donor-specific transplantation tolerance and treat autoimmune diseases. Blood 112, 3543–3553 (2008).
Seita, J. & Weissman, I. L. Hematopoietic stem cell: self-renewal versus differentiation. Wiley Interdiscip. Rev. Syst. Biol. Med. 2, 640–653 (2010).
Copelan, E. A. Hematopoietic stem-cell transplantation. N. Engl. J. Med. 354, 1813–1826 (2006).
Chabannon, C. et al. Hematopoietic stem cell transplantation in its 60s: a platform for cellular therapies. Sci. Transl. Med. 10, eaap9630 (2018).
Pinho, S. & Frenette, P. S. Haematopoietic stem cell activity and interactions with the niche. Nat. Rev. Mol. Cell Biol. 20, 303–320 (2019).
Spangrude, G. J., Heimfeld, S. & Weissman, I. L. Purification and characterization of mouse hematopoietic stem cells. Science 241, 58–62 (1988).
Visser, J. W., Bauman, J. G., Mulder, A. H., Eliason, J. F. & de Leeuw, A. M. Isolation of murine pluripotent hemopoietic stem cells. J. Exp. Med. 159, 1576–1590 (1984).
Kiel, M. J., Yilmaz, O. H., Iwashita, T., Terhorst, C. & Morrison, S. J. SLAM family receptors distinguish hematopoietic stem and progenitor cells and reveal endothelial niches for stem cells. Cell 121, 1109–1121 (2005).
Morita, Y., Ema, H. & Nakauchi, H. Heterogeneity and hierarchy within the most primitive hematopoietic stem cell compartment. J. Exp. Med. 207, 1173–1182 (2010).
Yamamoto, R. et al. Clonal analysis unveils self-renewing lineage-restricted progenitors generated directly from hematopoietic stem cells. Cell 154, 1112–1126 (2013).
Balazs, A. B., Fabian, A. J., Esmon, C. T. & Mulligan, R. C. Endothelial protein C receptor (CD201) explicitly identifies hematopoietic stem cells in murine bone marrow. Blood 107, 2317–2321 (2006).
Gazit, R. et al. Fgd5 identifies hematopoietic stem cells in the murine bone marrow. J. Exp. Med. 211, 1315–1331 (2014).
Chen, J. Y. et al. Hoxb5 marks long-term haematopoietic stem cells and reveals a homogenous perivascular niche. Nature 530, 223–227 (2016).
Kataoka, K. et al. Evi1 is essential for hematopoietic stem cell self-renewal, and its expression marks hematopoietic cells with long-term multilineage repopulating activity. J. Exp. Med. 208, 2403–2416 (2011).
Acar, M. et al. Deep imaging of bone marrow shows non-dividing stem cells are mainly perisinusoidal. Nature 526, 126–130 (2015).
Tajima, Y. et al. Continuous cell supply from Krt7-expressing hematopoietic stem cells during native hematopoiesis revealed by targeted in vivo gene transfer method. Sci. Rep. 7, 40684 (2017).
Notta, F. et al. Isolation of single human hematopoietic stem cells capable of long-term multilineage engraftment. Science 333, 218–221 (2011). This article describes the use of CD49f expression to purify human HSCs at high frequencies.
Wilson, N. K. et al. Combined single-cell functional and gene expression analysis resolves heterogeneity within stem cell populations. Cell Stem Cell 16, 712–724 (2015).
Notta, F. et al. Distinct routes of lineage development reshape the human blood hierarchy across ontogeny. Science 351, aab2116 (2016).
Wilkinson, A. C. et al. Long-term ex vivo haematopoietic-stem-cell expansion allows nonconditioned transplantation. Nature 571, 117–121 (2019). This article describes a long-term ex vivo expansion culture system for mouse HSCs using PVA.
Fares, I. et al. EPCR expression marks UM171-expanded CD34. Blood 129, 3344–3351 (2017).
Tomellini, E. et al. Integrin-α3 is a functional marker of ex vivo expanded human long-term hematopoietic stem cells. Cell Rep. 28, 1063–1073 (2019).
Till, J. E. & McCulloch, E. A. A direct measurement of the radiation sensitivity of normal mouse bone marrow cells. Radiat. Res. 14, 213–222 (1961).
Becker, A. J., McCulloch, E. A. & Till, J. E. Cytological demonstration of the clonal nature of spleen colonies derived from transplanted mouse marrow cells. Nature 197, 452–454 (1963).
Szilvassy, S. J., Humphries, R. K., Lansdorp, P. M., Eaves, A. C. & Eaves, C. J. Quantitative assay for totipotent reconstituting hematopoietic stem cells by a competitive repopulation strategy. Proc. Natl Acad. Sci. USA 87, 8736–8740 (1990).
Lemischka, I. R., Raulet, D. H. & Mulligan, R. C. Developmental potential and dynamic behavior of hematopoietic stem cells. Cell 45, 917–927 (1986).
Osawa, M., Hanada, K., Hamada, H. & Nakauchi, H. Long-term lymphohematopoietic reconstitution by a single CD34-low/negative hematopoietic stem cell. Science 273, 242–245 (1996). This article describes the use of single-cell transplantation assays to identify self-renewing multipotent HSCs within the mouse bone marrow.
Waterstrat, A., Liang, Y., Swiderski, C. F., Shelton, B. J. & Van Zant, G. Congenic interval of CD45/Ly-5 congenic mice contains multiple genes that may influence hematopoietic stem cell engraftment. Blood 115, 408–417 (2010).
Mercier, F. E., Sykes, D. B. & Scadden, D. T. Single targeted exon mutation creates a true congenic mouse for competitive hematopoietic stem cell transplantation: the C57BL/6-CD45.1STEM mouse. Stem Cell Rep. 6, 985–992 (2016).
Yamamoto, R. et al. Large-scale clonal analysis resolves aging of the mouse hematopoietic stem cell compartment. Cell Stem Cell 22, 600–607 (2018).
Carrelha, J. et al. Hierarchically related lineage-restricted fates of multipotent haematopoietic stem cells. Nature 554, 106–111 (2018). This article provides definitive evidence for platelet-restricted self-renewing stem cells.
Haas, S. et al. Inflammation-induced emergency megakaryopoiesis driven by hematopoietic stem cell-like megakaryocyte progenitors. Cell Stem Cell 17, 422–434 (2015).
Benveniste, P. et al. Intermediate-term hematopoietic stem cells with extended but time-limited reconstitution potential. Cell Stem Cell 6, 48–58 (2010).
Yamamoto, R., Wilkinson, A. C. & Nakauchi, H. Changing concepts in hematopoietic stem cells. Science 362, 895–896 (2018).
Ema, H., Takano, H., Sudo, K. & Nakauchi, H. In vitro self-renewal division of hematopoietic stem cells. J. Exp. Med. 192, 1281–1288 (2000).
Gerrits, A. et al. Cellular barcoding tool for clonal analysis in the hematopoietic system. Blood 115, 2610–2618 (2010).
Lu, R., Neff, N. F., Quake, S. R. & Weissman, I. L. Tracking single hematopoietic stem cells in vivo using high-throughput sequencing in conjunction with viral genetic barcoding. Nat. Biotechnol. 29, 928–933 (2011).
Naik, S. H. et al. Diverse and heritable lineage imprinting of early haematopoietic progenitors. Nature 496, 229–232 (2013).
Sun, J. et al. Clonal dynamics of native haematopoiesis. Nature 514, 322–327 (2014). This article describes the use of Sleeping Beauty transposon-based barcoding of HSCs and HPCs in vivo.
Busch, K. et al. Fundamental properties of unperturbed haematopoiesis from stem cells in vivo. Nature 518, 542–546 (2015).
Bradford, G. B., Williams, B., Rossi, R. & Bertoncello, I. Quiescence, cycling, and turnover in the primitive hematopoietic stem cell compartment. Exp. Hematol. 25, 445–453 (1997).
Cheshier, S. H., Morrison, S. J., Liao, X. & Weissman, I. L. In vivo proliferation and cell cycle kinetics of long-term self-renewing hematopoietic stem cells. Proc. Natl Acad. Sci. USA 96, 3120–3125 (1999).
Sudo, K., Ema, H., Morita, Y. & Nakauchi, H. Age-associated characteristics of murine hematopoietic stem cells. J. Exp. Med. 192, 1273–1280 (2000).
Wilson, A. et al. Hematopoietic stem cells reversibly switch from dormancy to self-renewal during homeostasis and repair. Cell 135, 1118–1129 (2008).
Yu, V. W. C. et al. Epigenetic memory underlies cell-autonomous heterogeneous behavior of hematopoietic stem cells. Cell 168, 944–945 (2017).
Ganuza, M. et al. The global clonal complexity of the murine blood system declines throughout life and after serial transplantation. Blood 133, 1927–1942 (2019).
Bernitz, J. M., Kim, H. S., MacArthur, B., Sieburg, H. & Moore, K. Hematopoietic stem cells count and remember self-renewal divisions. Cell 167, 1296–1309 (2016).
Pei, W. et al. Polylox barcoding reveals haematopoietic stem cell fates realized in vivo. Nature 548, 456–460 (2017). This article describes the development and validation of an in vivo Polylox barcode system that allows numerous HSCs to be barcoded and tracked during native haematopoiesis.
Frieda, K. L. et al. Synthetic recording and in situ readout of lineage information in single cells. Nature 541, 107–111 (2017).
Loeffler, D. et al. Asymmetric lysosome inheritance predicts activation of haematopoietic stem cells. Nature 573, 426–429 (2019).
Christodoulou, C. et al. Live-animal imaging of native haematopoietic stem and progenitor cells. Nature 578, 278–283 (2020).
Kamel-Reid, S. & Dick, J. E. Engraftment of immune-deficient mice with human hematopoietic stem cells. Science 242, 1706–1709 (1988).
Goyama, S., Wunderlich, M. & Mulloy, J. C. Xenograft models for normal and malignant stem cells. Blood 125, 2630–2640 (2015).
Yurino, A. et al. Enhanced reconstitution of human erythropoiesis and thrombopoiesis in an immunodeficient mouse model with KitWv mutations. Stem Cell Rep. 7, 425–438 (2016).
Rahmig, S. et al. Improved human erythropoiesis and platelet formation in humanized NSGW41 mice. Stem Cell Rep. 7, 591–601 (2016).
Miller, P. H. et al. Analysis of parameters that affect human hematopoietic cell outputs in mutant c-kit-immunodeficient mice. Exp. Hematol. 48, 41–49 (2017).
Takagi, S. et al. Membrane-bound human SCF/KL promotes in vivo human hematopoietic engraftment and myeloid differentiation. Blood 119, 2768–2777 (2012).
Biasco, L. et al. In vivo tracking of human hematopoiesis reveals patterns of clonal dynamics during early and steady-state reconstitution phases. Cell Stem Cell 19, 107–119 (2016).
Scala, S. et al. Dynamics of genetically engineered hematopoietic stem and progenitor cells after autologous transplantation in humans. Nat. Med. 24, 1683–1690 (2018).
Lee-Six, H. et al. Population dynamics of normal human blood inferred from somatic mutations. Nature 561, 473–478 (2018). This work uses whole-genome sequencing to identify somatic mutations that could be used to infer lineage relationships in human haematopoiesis.
Morrison, S. J., Hemmati, H. D., Wandycz, A. M. & Weissman, I. L. The purification and characterization of fetal liver hematopoietic stem cells. Proc. Natl Acad. Sci. USA 92, 10302–10306 (1995).
Ema, H. & Nakauchi, H. Expansion of hematopoietic stem cells in the developing liver of a mouse embryo. Blood 95, 2284–2288 (2000).
Gekas, C., Dieterlen-Lièvre, F., Orkin, S. H. & Mikkola, H. K. The placenta is a niche for hematopoietic stem cells. Dev. Cell 8, 365–375 (2005).
de Haan, G. & Lazare, S. S. Aging of hematopoietic stem cells. Blood 131, 479–487 (2018).
Ema, H. et al. Quantification of self-renewal capacity in single hematopoietic stem cells from normal and Lnk-deficient mice. Dev. Cell 8, 907–914 (2005).
Morrison, S. J. & Scadden, D. T. The bone marrow niche for haematopoietic stem cells. Nature 505, 327–334 (2014).
Crane, G. M., Jeffery, E. & Morrison, S. J. Adult haematopoietic stem cell niches. Nat. Rev. Immunol. 17, 573–590 (2017).
Zhang, C. C. & Lodish, H. F. Cytokines regulating hematopoietic stem cell function. Curr. Opin. Hematol. 15, 307–311 (2008).
Edling, C. E. & Hallberg, B. c-Kit — a hematopoietic cell essential receptor tyrosine kinase. Int. J. Biochem. Cell Biol. 39, 1995–1998 (2007).
Ding, L., Saunders, T. L., Enikolopov, G. & Morrison, S. J. Endothelial and perivascular cells maintain haematopoietic stem cells. Nature 481, 457–462 (2012).
de Graaf, C. A. & Metcalf, D. Thrombopoietin and hematopoietic stem cells. Cell Cycle 10, 1582–1589 (2011).
Decker, M., Leslie, J., Liu, Q. & Ding, L. Hepatic thrombopoietin is required for bone marrow hematopoietic stem cell maintenance. Science 360, 106–110 (2018).
Tadokoro, Y. et al. Spred1 safeguards hematopoietic homeostasis against diet-induced systemic stress. Cell Stem Cell 22, 713–725 (2018).
Seita, J. et al. Lnk negatively regulates self-renewal of hematopoietic stem cells by modifying thrombopoietin-mediated signal transduction. Proc. Natl Acad. Sci. USA 104, 2349–2354 (2007).
Takizawa, H. et al. Pathogen-induced TLR4-TRIF innate immune signaling in hematopoietic stem cells promotes proliferation but reduces competitive fitness. Cell Stem Cell 21, 225–240 (2017).
Pietras, E. M. et al. Chronic interleukin-1 exposure drives haematopoietic stem cells towards precocious myeloid differentiation at the expense of self-renewal. Nat. Cell Biol. 18, 607–618 (2016).
Takizawa, H. & Manz, M. G. Impact of inflammation on early hematopoiesis and the microenvironment. Int. J. Hematol. 106, 27–33 (2017).
Pietras, E. M. Inflammation: a key regulator of hematopoietic stem cell fate in health and disease. Blood 130, 1693–1698 (2017).
Wilkinson, A. C. & Yamazaki, S. The hematopoietic stem cell diet. Int. J. Hematol. 107, 634–641 (2018).
Cheng, C. W. et al. Prolonged fasting reduces IGF-1/PKA to promote hematopoietic-stem-cell-based regeneration and reverse immunosuppression. Cell Stem Cell 14, 810–823 (2014).
Lazare, S. et al. Lifelong dietary intervention does not affect hematopoietic stem cell function. Exp. Hematol. 53, 26–30 (2017).
Tang, D. et al. Dietary restriction improves repopulation but impairs lymphoid differentiation capacity of hematopoietic stem cells in early aging. J. Exp. Med. 213, 535–553 (2016).
Taya, Y. et al. Depleting dietary valine permits nonmyeloablative mouse hematopoietic stem cell transplantation. Science 354, 1152–1155 (2016).
Wilkinson, A. C., Morita, M., Nakauchi, H. & Yamazaki, S. Branched-chain amino acid depletion conditions bone marrow for hematopoietic stem cell transplantation avoiding amino acid imbalance-associated toxicity. Exp. Hematol. 63, 12–16 (2018).
Mantel, C. R. et al. Enhancing hematopoietic stem cell transplantation efficacy by mitigating oxygen shock. Cell 161, 1553–1565 (2015).
Bowie, M. B., Kent, D. G., Copley, M. R. & Eaves, C. J. Steel factor responsiveness regulates the high self-renewal phenotype of fetal hematopoietic stem cells. Blood 109, 5043–5048 (2007).
Prashad, S. L. et al. GPI-80 defines self-renewal ability in hematopoietic stem cells during human development. Cell Stem Cell 16, 80–87 (2015).
Chhabra, A. et al. Trophoblasts regulate the placental hematopoietic niche through PDGF-B signaling. Dev. Cell 22, 651–659 (2012).
Moran-Crusio, K. et al. Tet2 loss leads to increased hematopoietic stem cell self-renewal and myeloid transformation. Cancer Cell 20, 11–24 (2011).
Challen, G. A. et al. Dnmt3a is essential for hematopoietic stem cell differentiation. Nat. Genet. 44, 23–31 (2011).
Jeong, M. et al. Loss of Dnmt3a immortalizes hematopoietic stem cells in vivo. Cell. Rep. 23, 1–10 (2018).
Iwama, A. et al. Enhanced self-renewal of hematopoietic stem cells mediated by the polycomb gene product Bmi-1. Immunity 21, 843–851 (2004).
Park, I. K. et al. Bmi-1 is required for maintenance of adult self-renewing haematopoietic stem cells. Nature 423, 302–305 (2003).
Luis, T. C., Wilkinson, A. C., Beerman, I., Jaiswal, S. & Shlush, L. I. Biological implications of clonal hematopoiesis. Exp. Hematol. 77, 1–5 (2019).
Calvanese, V. et al. MLLT3 governs human haematopoietic stem-cell self-renewal and engraftment. Nature 576, 281–286 (2019).
Ito, K. et al. A PML–PPAR-δ pathway for fatty acid oxidation regulates hematopoietic stem cell maintenance. Nat. Med. 18, 1350–1358 (2012).
Ito, K. et al. Self-renewal of a purified Tie2+ hematopoietic stem cell population relies on mitochondrial clearance. Science 354, 1156–1160 (2016).
Ansó, E. et al. The mitochondrial respiratory chain is essential for haematopoietic stem cell function. Nat. Cell Biol. 19, 614–625 (2017).
Ho, T. T. et al. Autophagy maintains the metabolism and function of young and old stem cells. Nature 543, 205–210 (2017).
Ito, K. Hematopoietic stem cell fate through metabolic control. Exp. Hematol. 64, 1–11 (2018).
Agathocleous, M. et al. Ascorbate regulates haematopoietic stem cell function and leukaemogenesis. Nature 549, 476–481 (2017).
Cimmino, L. et al. Restoration of TET2 function blocks aberrant self-renewal and leukemia progression. Cell 170, 1079–1095 (2017).
Wilkinson, A. C. & Gottgens, B. Transcriptional regulation of haematopoietic stem cells. Adv. Exp. Med. Biol. 786, 187–212 (2013).
Kim, I., Saunders, T. L. & Morrison, S. J. Sox17 dependence distinguishes the transcriptional regulation of fetal from adult hematopoietic stem cells. Cell 130, 470–483 (2007).
He, S., Kim, I., Lim, M. S. & Morrison, S. J. Sox17 expression confers self-renewal potential and fetal stem cell characteristics upon adult hematopoietic progenitors. Genes. Dev. 25, 1613–1627 (2011).
Zhao, Y. et al. ATF4 plays a pivotal role in the development of functional hematopoietic stem cells in mouse fetal liver. Blood 126, 2383–2391 (2015).
Mochizuki-Kashio, M. et al. Dependency on the polycomb gene Ezh2 distinguishes fetal from adult hematopoietic stem cells. Blood 118, 6553–6561 (2011).
Copley, M. R. et al. The Lin28b-let-7-Hmga2 axis determines the higher self-renewal potential of fetal haematopoietic stem cells. Nat. Cell Biol. 15, 916–925 (2013).
Kumar, S. & Geiger, H. HSC niche biology and HSC expansion ex vivo. Trends Mol. Med. 23, 799–819 (2017).
Ku, H., Yonemura, Y., Kaushansky, K. & Ogawa, M. Thrombopoietin, the ligand for the Mpl receptor, synergizes with steel factor and other early acting cytokines in supporting proliferation of primitive hematopoietic progenitors of mice. Blood 87, 4544–4551 (1996).
Sitnicka, E. et al. The effect of thrombopoietin on the proliferation and differentiation of murine hematopoietic stem cells. Blood 87, 4998–5005 (1996).
Ramsfjell, V. et al. Thrombopoietin, but not erythropoietin, directly stimulates multilineage growth of primitive murine bone marrow progenitor cells in synergy with early acting cytokines: distinct interactions with the ligands for c-kit and FLT3. Blood 88, 4481–4492 (1996).
Miller, C. L. & Eaves, C. J. Expansion in vitro of adult murine hematopoietic stem cells with transplantable lympho-myeloid reconstituting ability. Proc. Natl Acad. Sci. USA 94, 13648–13653 (1997).
Himburg, H. A. et al. Pleiotrophin regulates the expansion and regeneration of hematopoietic stem cells. Nat. Med. 16, 475–482 (2010).
Ieyasu, A. et al. An all-recombinant protein-based culture system specifically identifies hematopoietic stem cell maintenance factors. Stem Cell Rep. 8, 500–508 (2017).
Butler, J. M. et al. Endothelial cells are essential for the self-renewal and repopulation of Notch-dependent hematopoietic stem cells. Cell Stem Cell 6, 251–264 (2010).
Nakahara, F. et al. Engineering a haematopoietic stem cell niche by revitalizing mesenchymal stromal cells. Nat. Cell Biol. 21, 560–567 (2019).
Bai, T. et al. Expansion of primitive human hematopoietic stem cells by culture in a zwitterionic hydrogel. Nat. Med. 25, 1566–1575 (2019). This article describes the novel use of zwitterionic hydrogel-based 3D cultures for expanding human HSCs ex vivo.
Umemoto, T. et al. Integrin-αvβ3 regulates thrombopoietin-mediated maintenance of hematopoietic stem cells. Blood 119, 83–94 (2012).
Antonchuk, J., Sauvageau, G. & Humphries, R. K. HOXB4-induced expansion of adult hematopoietic stem cells ex vivo. Cell 109, 39–45 (2002).
Miharada, K., Sigurdsson, V. & Karlsson, S. Dppa5 improves hematopoietic stem cell activity by reducing endoplasmic reticulum stress. Cell Rep. 7, 1381–1392 (2014).
Kunisato, A. et al. HES-1 preserves purified hematopoietic stem cells ex vivo and accumulates side population cells in vivo. Blood 101, 1777–1783 (2003).
Reya, T. et al. A role for Wnt signalling in self-renewal of haematopoietic stem cells. Nature 423, 409–414 (2003).
Boitano, A. E. et al. Aryl hydrocarbon receptor antagonists promote the expansion of human hematopoietic stem cells. Science 329, 1345–1348 (2010).
Fares, I. et al. Cord blood expansion. Pyrimidoindole derivatives are agonists of human hematopoietic stem cell self-renewal. Science 345, 1509–1512 (2014). This article describes the identification of the small molecule UM171, a potent human HSC self-renewal agonist.
Chagraoui, J. et al. UM171 induces a homeostatic inflammatory-detoxification response supporting human HSC self-renewal. PLoS ONE 14, e0224900 (2019).
Sun, H., Tsai, Y., Nowak, I., Liesveld, J. & Chen, Y. Eltrombopag, a thrombopoietin receptor agonist, enhances human umbilical cord blood hematopoietic stem/primitive progenitor cell expansion and promotes multi-lineage hematopoiesis. Stem Cell Res. 9, 77–86 (2012).
Nishino, T. et al. Ex vivo expansion of human hematopoietic stem cells by a small-molecule agonist of c-MPL. Exp. Hematol. 37, 1364–1377 (2009).
Csaszar, E. et al. Rapid expansion of human hematopoietic stem cells by automated control of inhibitory feedback signaling. Cell Stem Cell 10, 218–229 (2012).
Francis, G. L. Albumin and mammalian cell culture: implications for biotechnology applications. Cytotechnology 62, 1–16 (2010).
Wilkinson, A. C., Ishida, R., Nakauchi, H. & Yamazaki, S. Long-term ex vivo expansion of mouse hematopoietic stem cells. Nat. Protoc. 15, 628–648 (2020).
Nishimura, T. et al. Use of polyvinyl alcohol for chimeric antigen receptor T-cell expansion. Exp. Hematol. 80, 16–20 (2019).
Luchsinger, L. L. et al. Harnessing hematopoietic stem cell low intracellular calcium improves their maintenance in vitro. Cell Stem Cell 25, 225–240 (2019).
Umemoto, T., Hashimoto, M., Matsumura, T., Nakamura-Ishizu, A. & Suda, T. Ca2+-mitochondria axis drives cell division in hematopoietic stem cells. J. Exp. Med. 215, 2097–2113 (2018).
Kobayashi, H. et al. Environmental optimization enables maintenance of quiescent hematopoietic stem cells ex vivo. Cell Rep. 28, 145–158 (2019).
Morgan, R. A., Gray, D., Lomova, A. & Kohn, D. B. Hematopoietic stem cell gene therapy: progress and lessons learned. Cell Stem Cell 21, 574–590 (2017).
Negrin, R. S. Graft-versus-host disease versus graft-versus-leukemia. Hematol. Am. Soc. Hematol. Educ. Program. 2015, 225–230 (2015).
Broxmeyer, H. E. Enhancing the efficacy of engraftment of cord blood for hematopoietic cell transplantation. Transfus. Apher. Sci. 54, 364–372 (2016).
Kim, Y. J. & Broxmeyer, H. E. Immune regulatory cells in umbilical cord blood and their potential roles in transplantation tolerance. Crit. Rev. Oncol. Hematol. 79, 112–126 (2011).
Cohen, S. et al. Hematopoietic stem cell transplantation using single UM171-expanded cord blood: a single-arm, phase 1–2 safety and feasibility study. Lancet Haematol. 7, e134–e145 (2020). This recent phase I/II clinical trial report demonstrates the safety and feasibility of HSCT using ex vivo UM171-expanded umbilical cord blood HSCs.
Wagner, J. E. et al. Phase I/II trial of StemRegenin-1 expanded umbilical cord blood hematopoietic stem cells supports testing as a stand-alone graft. Cell Stem Cell 18, 144–155 (2016).
de Lima, M. et al. Cord-blood engraftment with ex vivo mesenchymal-cell coculture. N. Engl. J. Med. 367, 2305–2315 (2012).
Dever, D. P. & Porteus, M. H. The changing landscape of gene editing in hematopoietic stem cells: a step towards Cas9 clinical translation. Curr. Opin. Hematol. 24, 481–488 (2017).
Gundry, M. C. et al. Technical considerations for the use of CRISPR/Cas9 in hematology research. Exp. Hematol. 54, 4–11 (2017).
Bhattacharya, D., Rossi, D. J., Bryder, D. & Weissman, I. L. Purified hematopoietic stem cell engraftment of rare niches corrects severe lymphoid deficiencies without host conditioning. J. Exp. Med. 203, 73–85 (2006).
Shimoto, M., Sugiyama, T. & Nagasawa, T. Numerous niches for hematopoietic stem cells remain empty during homeostasis. Blood 129, 2124–2131 (2017).
Kitao, H. & Takata, M. Fanconi anemia: a disorder defective in the DNA damage response. Int. J. Hematol. 93, 417–424 (2011).
Evans, M. Discovering pluripotency: 30 years of mouse embryonic stem cells. Nat. Rev. Mol. Cell Biol. 12, 680–686 (2011).
Ivanovs, A. et al. Human haematopoietic stem cell development: from the embryo to the dish. Development 144, 2323–2337 (2017).
Wilkinson, A. C. Hope for hematological diseases. Science 367, 1206 (2020).
Laurenti, E. & Göttgens, B. From haematopoietic stem cells to complex differentiation landscapes. Nature 553, 418–426 (2018).
Haas, S., Trumpp, A. & Milsom, M. D. Causes and consequences of hematopoietic stem cell heterogeneity. Cell Stem Cell 22, 627–638 (2018).
Dzierzak, E. & Speck, N. A. Of lineage and legacy: the development of mammalian hematopoietic stem cells. Nat. Immunol. 9, 129–136 (2008).
de Bruijn, M. F., Speck, N. A., Peeters, M. C. & Dzierzak, E. Definitive hematopoietic stem cells first develop within the major arterial regions of the mouse embryo. EMBO J. 19, 2465–2474 (2000).
Ivanovs, A. et al. Highly potent human hematopoietic stem cells first emerge in the intraembryonic aorta-gonad-mesonephros region. J. Exp. Med. 208, 2417–2427 (2011).
Ottersbach, K. & Dzierzak, E. The murine placenta contains hematopoietic stem cells within the vascular labyrinth region. Dev. Cell 8, 377–387 (2005).
Kumaravelu, P. et al. Quantitative developmental anatomy of definitive haematopoietic stem cells/long-term repopulating units (HSC/RUs): role of the aorta-gonad-mesonephros (AGM) region and the yolk sac in colonisation of the mouse embryonic liver. Development 129, 4891–4899 (2002).
A.C.W. is supported by the Leukemia & Lymphoma Society (grant 3385-19) and the US National Institutes of Health (grant K99HL150218). K.J.I. is supported by the US National Science Foundation (grant 2018261442). H.N. is supported by the California Institute for Regenerative Medicine (grants LA1_C12-06917 and DISC1-10555), the US National Institutes of Health (grants R01DK116944, R01HL147124 and R21AG061487), JSPS KAKENHI Grant-in-Aid for Scientific Research, AMED Advanced Research and Development Programs for Medical Innovation (LEAP), and the Virginia and D.K. Ludwig Fund for Cancer Research. The article content is solely the responsibility of the authors and does not necessarily represent the official views of the US National Institutes of Health.
H.N. declares that he is a co-founder and shareholder in ReproCELL, Megakaryon and Century Therapeutics. The other authors declare no competing interests.
Peer review information
Nature Reviews Genetics thanks G. Sauvageau, P. Frenette and the other, anonymous, reviewer(s) for their contribution to the peer review of this work.
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
About this article
Cite this article
Wilkinson, A.C., Igarashi, K.J. & Nakauchi, H. Haematopoietic stem cell self-renewal in vivo and ex vivo. Nat Rev Genet 21, 541–554 (2020). https://doi.org/10.1038/s41576-020-0241-0
Stem Cell Research & Therapy (2020)