In vitro conversion of adult murine endothelial cells to hematopoietic stem cells.

2017 ◽  
Author(s):  
Raphael Lis ◽  
Raphael Lis ◽  
Charles C. Karrasch ◽  
Shahin Rafii
Keyword(s):  
Stem Cells ◽  
Endothelial Cells ◽  
Hematopoietic Stem Cells ◽  
Hematopoietic Stem

In vitro conversion of adult murine endothelial cells to hematopoietic stem cells

2018 ◽  
Vol 13 (12) ◽  
pp. 2758-2780 ◽  
Author(s):  
José Gabriel Barcia Durán ◽  
Raphaël Lis ◽  
Tyler M. Lu ◽  
Shahin Rafii
Keyword(s):  
Stem Cells ◽  
Endothelial Cells ◽  
Hematopoietic Stem Cells ◽  
Hematopoietic Stem

scholarly journals Distinct Transcriptional Signatures Distinguish the Emergence of Multipotent Progenitors and Hematopoietic Stem Cells from Endothelial Precursors in the Murine Embryo

Blood ◽  
2020 ◽  
Vol 136 (Supplement 1) ◽  
pp. 7-8
Author(s):  
Tessa Dignum ◽  
Barbara Varnum-Finney ◽  
Sanjay Srivatsan ◽  
Stacey Dozono ◽  
Olivia Waltner ◽  
...  
Keyword(s):  
Stem Cells ◽  
Endothelial Cells ◽  
Hematopoietic Stem Cells ◽  
Surface Markers ◽  
Publicly Traded ◽  
Self Renewal ◽  
Hematopoietic Stem ◽  
Multipotent Progenitors ◽  
Endothelial Precursors

During embryonic development, blood cells emerge from hemogenic endothelium (HE), producing waves of hematopoietic progenitors prior to the emergence of rare hematopoietic stem cells (HSCs), which have the unique ability to self-renew and generate all cell types of the adult hematopoietic system. HSCs have significant potential for use in cellular therapies and disease modeling. However, efforts to generate HSCs in vitro from pluripotent stem cells (PSCs) have been limited by an incomplete understanding of the unique phenotypic markers and transcriptional programs that distinguish HE with HSC potential. Previous studies have demonstrated that yolk sac-derived erythromyeloid progenitors and HSCs originate from distinct populations of HE. However, it is not known whether the earliest lymphoid-competent progenitors, multipotent progenitors, and HSCs originate from HE with common phenotypic and transcriptional properties. To investigate this, we combined index sorting of single hemogenic precursors with stromal co-culture that enables simultaneous detection of HSC and multilineage hematopoietic potential, to functionally validate surface markers that may distinguish hemogenic precursors with different hematopoietic fates. We previously found that the co-expression of two markers, CD61 and EPCR, identifies a subset of VE-Cadherin+ endothelial cells from the mouse P-Sp/AGM region (para-aortic splanchnopleura/aorta-gonad-mesonephros, where the first HSCs are generated from HE between E9 and E11 in development) enriched phenotypically for arterial endothelial surface markers (e.g. Dll4, CD44) and functionally for hemogenic precursors with HSC potential. However, this population remains heterogeneous, containing clonal hemogenic precursors with the potential for HSC as well as multilineage progenitor-restricted fates. Here, we report that expression of arterial marker CXCR4 further enriched for functional HSC potential in hemogenic precursors in the P-Sp/AGM between E9 and E10, when the first clonal HSC precursors are detected at rare frequency. In contrast, we detected more abundant clonal HE with multilineage hematopoietic potential (producing lymphoid, erythroid, and myeloid progeny in vitro but lacking HSC potential) at the same stage, which are distinguished by comparatively lower CXCR4 expression. To investigate transcriptional differences between HE populations differentially expressing CXCR4, we performed single-cell RNA sequencing of E9 P-Sp-derived VE-Cadherin+CD61+EPCR+ cells. Using an unbiased gene module analysis based on graph autocorrelation in the Monocle 3 platform to identify genes that co-vary over pseudotime, we found that Cxcr4 is uniquely expressed in a subset of cells simultaneously enriched for arterial-specific genes (including Dll4, Efnb2, Hey2, Sox17, Cd44) and genes with established roles in HSC maintenance and self-renewal (including Mecom, Cdkn1c, H19, Txnip, Kmt2a). Conversely, expression of these genes is decreased in cells undergoing the endothelial to hematopoietic transition at this stage based on pseudotemporal ordering, concomitant with increasing expression of hematopoietic-specifying transcription factors Runx1 and Gfi1, and other genes associated with definitive hematopoiesis (egs. Myb, Kit, Hlf, Gata2, Mpl, Lyl1). We also examined the aggregate expression of established HSC-specific signature genes from published data sets across pseudotime, and found that they exhibit similar expression dynamics to that of Cxcr4 and Dll4, reaching peak expression prior to the initiation of Runx1 and Gfi1 expression. Altogether, our studies support a model in which the initial populations of multipotent progenitors and HSCs emerge independently from HE in the P-Sp/AGM. Furthermore, our findings suggest that HE with HSC competence is uniquely defined by co-expression of arterial endothelial genes and genes encoding HSC self-renewal factors, providing insight into the earliest transcriptional programs that must be coordinated to drive HSC fate from endothelial precursors. Future studies will focus on identifying the signal pathways whose integration promotes expression of these HSC-defining transcriptional programs in endothelial cells, with the goal of advancing methods for HSC generation in vitro. Disclosures Bernstein: Lyell Immunopharma: Current equity holder in publicly-traded company, Other: Laboratory Support; Deverra Therapeutics: Current equity holder in publicly-traded company.

Crosstalk between Murine Liver Sinusoidal Endothelial Cells (LSEC) and Hematopoietic Stem Cells during In Vitro Hematopoiesis.

Blood ◽  
2006 ◽  
Vol 108 (11) ◽  
pp. 4245-4245
Author(s):  
Jose E. Cardier ◽  
Gonzalo Luna
Keyword(s):  
Stem Cells ◽  
Endothelial Cells ◽  
Hematopoietic Stem Cells ◽  
Liver Sinusoidal Endothelial Cells ◽  
Sinusoidal Endothelial Cells ◽  
Hematopoietic Stem ◽  
Gm Csf ◽  
Hematopoietic Microenvironment ◽  
Murine Liver

Abstract Crosstalk between murine liver sinusoidal endothelial cells (LSEC) and hematopoietic stem cells during in vitro hematopoiesis. G. Luna, J. E. Cardier. Laboratorio de Patologia Celular y Molecular, Centro de Medicina Experimental, Instituto Venezolano de Investigaciones Científicas (IVIC). Apartado 21827, Caracas, Venezuela We have showed that at the liver there is a specific hematopoietic microenvironment, constituted by the liver sinusoid endothelial cells (LSEC), which are capable to support, in vitro and in vivo, not only the proliferation but also the differentiation and survival of hematopoietic stem cells (HSC). The ability of LSEC to support hematopoiesis could be related to specific hematopoietic molecules expressed by these cells. In this study, we investigate the expression of some cell adhesion molecules (VCAM-1, ICAM-1, and the integrin a4b1 ), and early acting hematopoietic cytokines (IL-6 and GM-CSF), on LSEC cocultured with HSC. The expression of VCAM-1, ICAM-1, α4β1, IL6 and GM-SCF was increased on LSEC cocultured with HSC. In contrast, a significant decrease in the expression of IL-6 and GM-CSF in the HSC derived from the same cocultures was observed. The blockade of VCAM-1 on LSEC reduced significantly the adhesion of HSC to LSEC monolayers, suggesting that this molecule is involved in the binding of HSC to LSEC microenvironments. There were not changes in the expression of the molecules evaluated when the LSEC and HSC were co-cultured in non-contact conditions, suggesting that soluble factors do not participate in regulating the expression of these molecules. Our data shows that during in vitro hematopoiesis, LSEC are activated to express molecules associated with the hematopoiesis process. LSEC activation is regulated by the contact between these cells and HSC. By expressing critical hematopoietic microenvironment molecules, LSEC may regulate the proliferation and differentiation of HSC, during liver extramedullary hematopoiesis.

Strategies to Protect Hematopoietic Stem Cells from Culture-Induced Stress Conditions

2020 ◽  
Vol 15 ◽  
Author(s):  
Fatima Aerts-Kaya
Keyword(s):  
Stem Cells ◽  
Hematopoietic Stem Cells ◽  
Ex Vivo ◽  
Stress Conditions ◽  
Stress Factors ◽  
Hematopoietic Stem ◽  
Induced Stress

: In contrast to their almost unlimited potential for expansion in vivo and despite years of dedicated research and optimization of expansion protocols, the expansion of Hematopoietic Stem Cells (HSCs) in vitro remains remarkably limited. Increased understanding of the mechanisms that are involved in maintenance, expansion and differentiation of HSCs will enable the development of better protocols for expansion of HSCs. This will allow procurement of HSCs with long-term engraftment potential and a better understanding of the effects of the external influences in and on the hematopoietic niche that may affect HSC function. During collection and culture of HSCs, the cells are exposed to suboptimal conditions that may induce different levels of stress and ultimately affect their self-renewal, differentiation and long-term engraftment potential. Some of these stress factors include normoxia, oxidative stress, extra-physiologic oxygen shock/stress (EPHOSS), endoplasmic reticulum (ER) stress, replicative stress, and stress related to DNA damage. Coping with these stress factors may help reduce the negative effects of cell culture on HSC potential, provide a better understanding of the true impact of certain treatments in the absence of confounding stress factors. This may facilitate the development of better ex vivo expansion protocols of HSCs with long-term engraftment potential without induction of stem cell exhaustion by cellular senescence or loss of cell viability. This review summarizes some of available strategies that may be used to protect HSCs from culture-induced stress conditions.

EXPANSION OF HEMATOPOIETIC STEM CELLS IN VITRO AS A MODEL SYSTEM FOR HUMAN TISSUE ENGINEERING

2000 ◽  
Vol 31 (3) ◽  
pp. 499-509 ◽  
Author(s):  
Joel S. Greenberger ◽  
Julie P. Goff ◽  
Jason Bush ◽  
Alfred Bahnson ◽  
Douglas Koebler ◽  
...  
Keyword(s):  
Tissue Engineering ◽  
Stem Cells ◽  
Hematopoietic Stem Cells ◽  
Human Tissue ◽  
Model System ◽  
Hematopoietic Stem

Generation of Hematopoietic Stem Cells from Purified Embryonic Endothelial Cells by a Simple and Efficient Strategy

2013 ◽  
Vol 40 (11) ◽  
pp. 557-563 ◽  
Author(s):  
Zhuan Li ◽  
Fan Zhou ◽  
Dongbo Chen ◽  
Wenyan He ◽  
Yanli Ni ◽  
...  
Keyword(s):  
Stem Cells ◽  
Endothelial Cells ◽  
Hematopoietic Stem Cells ◽  
Hematopoietic Stem ◽  
Efficient Strategy

3040 – IN VITRO RECAPITULATION OF MURINE T CELL DEVELOPMENT FROM SINGLE HEMATOPOIETIC STEM CELLS

2020 ◽  
Vol 88 ◽  
pp. S51
Author(s):  
Victoria Sun ◽  
Amelie Montel-Hagen ◽  
David Casero ◽  
Steven Tsai ◽  
Alexandre Zampieri ◽  
...  
Keyword(s):  
Stem Cells ◽  
T Cell ◽  
Hematopoietic Stem Cells ◽  
T Cell Development ◽  
Cell Development ◽  
Hematopoietic Stem
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