Human haematopoietic stem cell development: from the embryo to the dish
TL;DR: The extent to which in vitro differentiation of human pluripotent stem cells recapitulates bona fide human developmental haematopoiesis is discussed, and some future directions in the field are outlined.
Abstract: Haematopoietic stem cells (HSCs) emerge during embryogenesis and give rise to the adult haematopoietic system. Understanding how early haematopoietic development occurs is of fundamental importance for basic biology and medical sciences, but our knowledge is still limited compared with what we know of adult HSCs and their microenvironment. This is particularly true for human haematopoiesis, and is reflected in our current inability to recapitulate the development of HSCs from pluripotent stem cells in vitro In this Review, we discuss what is known of human haematopoietic development: the anatomical sites at which it occurs, the different temporal waves of haematopoiesis, the emergence of the first HSCs and the signalling landscape of the haematopoietic niche. We also discuss the extent to which in vitro differentiation of human pluripotent stem cells recapitulates bona fide human developmental haematopoiesis, and outline some future directions in the field.
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TL;DR: These evolving views of haematopoiesis have broad implications for the understanding of the functions of adult stem cells, as well as the development of new therapies for malignant and non-malignant haem atopoietic diseases.
Abstract: The development of mature blood cells from haematopoietic stem cells has long served as a model for stem-cell research, with the haematopoietic differentiation tree being widely used as a model for the maintenance of hierarchically organized tissues. Recent results and new technologies have challenged the demarcations between stem and progenitor cell populations, the timing of cell-fate choices and the contribution of stem and multipotent progenitor cells to the maintenance of steady-state blood production. These evolving views of haematopoiesis have broad implications for our understanding of the functions of adult stem cells, as well as the development of new therapies for malignant and non-malignant haematopoietic diseases.
491 citations
TL;DR: A shift in the haemopoietic composition of fetal liver during gestation away from being predominantly erythroid, accompanied by a parallel change in differentiation potential of HSC/MPPs is demonstrated, which is validated to produce an integrated map of fetal liver haematopoiesis.
Abstract: Definitive haematopoiesis in the fetal liver supports self-renewal and differentiation of haematopoietic stem cells/multipotent progenitors (HSC/MPPs) but remains poorly defined in humans. Using single cell transcriptome profiling of ~140,000 liver and ~74,000 skin, kidney and yolk sac cells, we identify the repertoire of human blood and immune cells during development. We infer differentiation trajectories from HSC/MPPs and evaluate the impact of tissue microenvironment on blood and immune cell development. We reveal physiological erythropoiesis in fetal skin and the presence of mast cells, NK and ILC precursors in the yolk sac. We demonstrate a shift in fetal liver haematopoietic composition during gestation away from being erythroid-predominant, accompanied by a parallel change in HSC/MPP differentiation potential, which we functionally validate. Our integrated map of fetal liver haematopoiesis provides a blueprint for the study of paediatric blood and immune disorders, and a valuable reference for harnessing the therapeutic potential of HSC/MPPs.
356 citations
TL;DR: D deterministic barcoding in tissue for spatial omics sequencing (DBiT-seq) is presented for co-mapping of mRNAs and proteins in a formaldehyde-fixed tissue slide via next-generation sequencing (NGS) and can be adopted by researchers with no experience in microfluidics.
327 citations
Journal Article•
TL;DR: It is shown that it is feasible to differentiate and mature human embryonic stem cells (hESCs) into functional oxygen-carrying erythrocytes on a large scale (10(10)-10(11) cells/6-well plate hESCs).
271 citations
TL;DR: The data represent a comprehensive characterization of the spatiotemporal dynamics of early macrophage development during human embryogenesis, providing a reference for future studies of the development and function of human TRMs.
Abstract: Macrophages are the first cells of the nascent immune system to emerge during embryonic development. In mice, embryonic macrophages infiltrate developing organs, where they differentiate symbiotically into tissue-resident macrophages (TRMs)1. However, our understanding of the origins and specialization of macrophages in human embryos is limited. Here we isolated CD45+ haematopoietic cells from human embryos at Carnegie stages 11 to 23 and subjected them to transcriptomic profiling by single-cell RNA sequencing, followed by functional characterization of a population of CD45+CD34+CD44+ yolk sac-derived myeloid-biased progenitors (YSMPs) by single-cell culture. We also mapped macrophage heterogeneity across multiple anatomical sites and identified diverse subsets, including various types of embryonic TRM (in the head, liver, lung and skin). We further traced the specification trajectories of TRMs from either yolk sac-derived primitive macrophages or YSMP-derived embryonic liver monocytes using both transcriptomic and developmental staging information, with a focus on microglia. Finally, we evaluated the molecular similarities between embryonic TRMs and their adult counterparts. Our data represent a comprehensive characterization of the spatiotemporal dynamics of early macrophage development during human embryogenesis, providing a reference for future studies of the development and function of human TRMs. Single-cell RNA sequencing of haematopoietic cells from human embryos at different developmental stages sheds light on the development and specification of macrophages in different tissues.
231 citations
References
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TL;DR: Induction of pluripotent stem cells from mouse embryonic or adult fibroblasts by introducing four factors, Oct3/4, Sox2, c-Myc, and Klf4, under ES cell culture conditions is demonstrated and iPS cells, designated iPS, exhibit the morphology and growth properties of ES cells and express ES cell marker genes.
23,959 citations
"Human haematopoietic stem cell deve..." refers background in this paper
...…and patterning In the mammalian embryo, ingression of cells through the primitive streak during gastrulation generates mesoderm, which colonises the yolk sac (extra-embryonic mesoderm) and intra-embryonic sites including paraxial, intermediate and lateral plate mesoderm (Tam and Behringer, 1997)....
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TL;DR: In this article, the major open reading frame encoded by this cDNA contains a short protein segment similar to a sequence present in the myc protein family, and the expression of one of these cDNAs transfected into C3H10T1/2 fibroblasts, where it is not normally expressed, is sufficient to convert them to stable myoblasts.
3,162 citations
"Human haematopoietic stem cell deve..." refers background in this paper
...Although enforced gene expression has been shown to be a powerful tool for altering cell fate for several decades (Davis et al., 1987; Kulessa et al., 1995), the real boost to the generation of blood cells using this approach was precipitated by reprogramming studies in which fibroblasts were reverted back to pluripotency using four transcription factors (Takahashi and Yamanaka, 2006; Pereira et al....
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...…and reprogramming to blood Although enforced gene expression has been shown to be a powerful tool for altering cell fate for several decades (Davis et al., 1987; Kulessa et al., 1995), the real boost to the generation of blood cells using this approach was precipitated by reprogramming…...
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TL;DR: A novel in vitro organ culture system is presented demonstrating that, at day 10 in gestation, hematopoietic stem cells initiate autonomously and exclusively within the aorta-gonad-mesonephros (AGM) region, suggesting that the AGM region is the source of the definitive adult hematosynthesis system, which subsequently colonizes the liver.
1,498 citations
"Human haematopoietic stem cell deve..." refers background in this paper
...The third wave, which includes self-renewing haematopoietic stem cells (HSCs) that give rise to the permanent adult haematopoietic system, emerges later inside the body of the embryo (intra-embryonic haematopoiesis) in the aorta-gonadmesonephros (AGM) region, which is evolutionarily conserved in many vertebrates (Medvinsky et al., 1993; Medvinsky and Dzierzak, 1996)....
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...…(HSCs) that give rise to the permanent adult haematopoietic system, emerges later inside the body of the embryo (intra-embryonic haematopoiesis) in the aorta-gonadmesonephros (AGM) region, which is evolutionarily conserved in many vertebrates (Medvinsky et al., 1993; Medvinsky and Dzierzak, 1996)....
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...In the mouse, this issue was addressed by pre-culturing embryonic tissues as explants to allow their further development ex vivo before assessing their haematopoietic potential (Cumano et al., 1996; Medvinsky and Dzierzak, 1996)....
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TL;DR: Experimental data are presented showing that human fetal liver hematopoietic cells, human fetal thymus, and human fetal lymph node support the differentiation of mature human T cells and B cells after engraftment into mice with genetically determined severe combined immunodeficiency.
Abstract: The study of human hematopoietic cells and the human immune system is hampered by the lack of a suitable experimental model. Experimental data are presented showing that human fetal liver hematopoietic cells, human fetal thymus, and human fetal lymph node support the differentiation of mature human T cells and B cells after engraftment into mice with genetically determined severe combined immunodeficiency. The resultant SCID-hu mice are found to have a transient wave of human CD4+ and CD8+ T cells and human IgG (immunoglobulin G) in the peripheral circulation. The functional status of the human immune system within this mouse model is not yet known.
1,470 citations
"Human haematopoietic stem cell deve..." refers background in this paper
...At about the same time, severe combined immunodeficiency (SCID) mice, which could accept human haematopoietic grafts, were generated (McCune et al., 1988)....
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Journal Article•
TL;DR: The multiple defects in innate and adaptive immunity unique to the NOD/LtSz-scid/scid mouse provide an excellent in vivo environment for reconstitution with human hematopoietic cells.
Abstract: The scid mutation was backcrossed ten generations onto the NOD/Lt strain background, resulting in an immunodeficient stock (NOD/LtSz-scid/scid) with multiple defects in adaptive as well as nonadaptive immunologic function. NOD/LtSz-scid/scid mice lack functional lymphoid cells and show little or no serum Ig with age. Although NOD/(Lt-)+/+ mice develop T cell-mediated autoimmune, insulin-dependent diabetes mellitus, NOD/LtSz-scid/scid mice are both insulitis- and diabetes-free throughout life. However, because of a high incidence of thymic lymphomas, the mean lifespan of this congenic stock is only 8.5 mo under specific pathogen-free conditions. After i.v. injection of human CEM T-lymphoblastoid cells, splenic engraftment of these cells was fourfold greater in NOD/LtSz-scid/scid mice than in C.B17/Sz-scid/scid mice. Although C.B-17Sz-scid/scid mice exhibit robust NK cell activity, this activity is markedly reduced in both NOD/(Lt-)+/+ and NOD/LtSz-scid/scid mice. Presence of a functionally less mature macrophage population in NOD/LtSz-scid/scid vs C.B-17Sz-scid/scid mice is indicated by persistence in the former of the NOD/Lt strain-specific defect in LPS-stimulated IL-1 secretion by marrow-derived macrophages. Although C.B-17Sz-scid/scid and C57BL/6Sz-scid/scid mice have elevated serum hemolytic complement activity compared with their respective +/+ controls, both NOD/(LtSz-)+/+ and NOD/LtSz-scid/scid mice lack this activity. Age-dependent increases in serum Ig levels (> 1 micrograms/ml) were observed in only 2 of 30 NOD/LtSz-scid/scid mice vs 21 of 29 C.B-17/Sz-scid/scid animals. The multiple defects in innate and adaptive immunity unique to the NOD/LtSz-scid/scid mouse provide an excellent in vivo environment for reconstitution with human hematopoietic cells.
1,221 citations
"Human haematopoietic stem cell deve..." refers background in this paper
...For a long time, NOD/SCID recipient mice (Shultz et al., 1995) were widely used to study human haematopoiesis in vivo....
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