Transforming growth factor β1 accelerates and enhances in vitro red blood cell formation from hematopoietic stem cells by stimulating mitophagy

Pluripotent hematopoietic stem cells (PHSC) are very rare cells whose functional capabilities can only be analyzed indirectly. For a better understanding and possible manipulation of mechanisms that regulate self-renewal and commitment to differentiation of PHSC, it is necessary to purify these cells and to develop assays for their growth in vitro. In the present study, a rapid and simple, widely applicable procedure to highly purify day 14 spleen colony-forming cells (day 14 CFU-S) is described. Low density bone marrow cells (rho less than or equal to 1.078 g/cm3) were enriched by two successive light-activated cell sorting procedures. In the first sort, cells within a predetermined light scatter (blast cell) window that are wheat germ agglutinin/Texas Red (WGA/TxR) positive and mAb 15-1.4.1/fluorescein isothiocyanate negative (granulocyte-monocyte marker) were selected. In the second sort, cells were selected on the basis of retention of the supravital dye rhodamine 123 (Rh123). Cells that take up little Rh123 (Rh123 dull cells) and those that take up more Rh123 (Rh123 bright cells) were 237-fold and 132-fold enriched, respectively, for day 14 CFU-S. Both Rh123 fractions were cultured for various time periods in vitro in the presence of mast cell growth factor (MGF), with or without interleukin 3 (IL-3) or IL-1 alpha. Both Rh123 fractions proliferated in response to MGF alone as determined by a [3H]TdR assay or by counting nucleated cells present in the cultures over time. MGF also acted synergistically with both IL-3 and IL-1 alpha to promote stem cell proliferation. Stimulation of both Rh123 fractions with MGF alone did not result in a net increase of day 14 CFU-S. Stimulation with MGF + IL-3 or MGF + IL-alpha resulted in a 4.4- or 2.6-fold increase of day 14 CFU-S in the Rh123 dull fraction, and an 11.6-fold or 2.6-fold increase of day 14 CFU-S in the Rh123 bright fraction, respectively. The data presented in this paper indicate that in vitro MGF acts on primitive hematopoietic stem cells by itself and also is a potent synergistic factor in combination with IL-3 or IL-1 alpha.

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Osteogenic protein-1 with transforming growth factor-β1: potent inducer of chondrogenesis of synovial mesenchymal stem cells in vitro

Journal of Orthopaedic Science ◽

10.1007/s00776-007-1176-4 ◽

2007 ◽

Vol 12 (6) ◽

pp. 555-561 ◽

Cited By ~ 28

Author(s):

Chikara Miyamoto ◽

Tomoko Matsumoto ◽

Koichiro Sakimura ◽

Hiroyuki Shindo

Keyword(s):

Stem Cells ◽

Mesenchymal Stem Cells ◽

Growth Factor ◽

Transforming Growth Factor ◽

Transforming Growth Factor Β1 ◽

Potent Inducer ◽

Osteogenic Protein

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Smad4 is critical for self-renewal of hematopoietic stem cells

Journal of Experimental Medicine ◽

10.1084/jem.20060465 ◽

2007 ◽

Vol 204 (3) ◽

pp. 467-474 ◽

Cited By ~ 88

Author(s):

Göran Karlsson ◽

Ulrika Blank ◽

Jennifer L. Moody ◽

Mats Ehinger ◽

Sofie Singbrant ◽

...

Keyword(s):

Stem Cells ◽

Hematopoietic Stem Cells ◽

Transforming Growth Factor ◽

Transforming Growth Factor Β ◽

In Vitro Proliferation ◽

Self Renewal ◽

Hematopoietic Stem ◽

Early Embryonic Lethality

Members of the transforming growth factor β (TGF-β) superfamily of growth factors have been shown to regulate the in vitro proliferation and maintenance of hematopoietic stem cells (HSCs). Working at a common level of convergence for all TGF-β superfamily signals, Smad4 is key in orchestrating these effects. The role of Smad4 in HSC function has remained elusive because of the early embryonic lethality of the conventional knockout. We clarify its role by using an inducible model of Smad4 deletion coupled with transplantation experiments. Remarkably, systemic induction of Smad4 deletion through activation of MxCre was incompatible with survival 4 wk after induction because of anemia and histopathological changes in the colonic mucosa. Isolation of Smad4 deletion to the hematopoietic system via several transplantation approaches demonstrated a role for Smad4 in the maintenance of HSC self-renewal and reconstituting capacity, leaving homing potential, viability, and differentiation intact. Furthermore, the observed down-regulation of notch1 and c-myc in Smad4−/− primitive cells places Smad4 within a network of genes involved in the regulation HSC renewal.

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Transforming Growth Factor β, Pleiotropic Regulator of Hematopoietic Stem Cells: Potential Physiological and Clinical Relevance

International Journal of Hematology ◽

10.1007/bf02982545 ◽

2001 ◽

Vol 74 (1) ◽

pp. 18-25 ◽

Cited By ~ 21

Author(s):

Francis W. Ruscetti ◽

Stephen H. Bartelmez

Keyword(s):

Stem Cells ◽

Growth Factor ◽

Hematopoietic Stem Cells ◽

Clinical Relevance ◽

Transforming Growth Factor ◽

Transforming Growth Factor Β ◽

Hematopoietic Stem ◽

Pleiotropic Regulator

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Understanding the Journey of Human Hematopoietic Stem Cell Development

Stem Cells International ◽

10.1155/2019/2141475 ◽

2019 ◽

Vol 2019 ◽

pp. 1-13 ◽

Cited By ~ 6

Author(s):

Akhilesh Kumar ◽

Saritha S. D’Souza ◽

Abir S. Thakur

Keyword(s):

Stem Cells ◽

Blood Cell ◽

Hematopoietic Stem Cells ◽

Human Blood ◽

Molecular Mechanisms ◽

Inflammatory Responses ◽

Current Knowledge ◽

Hematopoietic Stem ◽

Hematopoietic Development

Hematopoietic stem cells (HSCs) surface during embryogenesis leading to the genesis of the hematopoietic system, which is vital for immune function, homeostasis balance, and inflammatory responses in the human body. Hematopoiesis is the process of blood cell formation, which initiates from hematopoietic stem/progenitor cells (HSPCs) and is responsible for the generation of all adult blood cells. With their self-renewing and pluripotent properties, human pluripotent stem cells (hPSCs) provide an unprecedented opportunity to createin vitromodels of differentiation that will revolutionize our understanding of human development, especially of the human blood system. The utilization of hPSCs provides newfound approaches for studying the origins of human blood cell diseases and generating progenitor populations for cell-based treatments. Current shortages in our knowledge of adult HSCs and the molecular mechanisms that control hematopoietic development in physiological and pathological conditions can be resolved with better understanding of the regulatory networks involved in hematopoiesis, their impact on gene expression, and further enhance our ability to develop novel strategies of clinical importance. In this review, we delve into the recent advances in the understanding of the various cellular and molecular pathways that lead to blood development from hPSCs and examine the current knowledge of human hematopoietic development. We also review howin vitrodifferentiation of hPSCs can undergo hematopoietic transition and specification, including major subtypes, and consider techniques and protocols that facilitate the generation of hematopoietic stem cells.

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