Article

Efficient enucleation of erythroblasts differentiated in vitro from hematopoietic stem and progenitor cells

Cell Engineering Division, RIKEN BioResource Center, Koyadai 3-1-1, Tsukuba, Ibaraki 305-0074, Japan.
Nature Biotechnology (Impact Factor: 39.08). 11/2006; 24(10):1255-6. DOI: 10.1038/nbt1245
Source: PubMed

ABSTRACT Erythroblast enucleation is thought to be largely dependent on signals mediated by other cells, such as macrophages. In an attempt to improve the in vitro production of red blood cells (RBCs) from immature hematopoietic progenitor cells, we have developed a method to produce enucleated RBCs efficiently in the absence of feeder cells. Our method may represent an efficient way to produce transfusable RBCs on a large scale from hematopoietic progenitors.

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Available from: Takashi Hiroyama, Feb 27, 2014
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    • "The most successful of these to date showed enucleation levels of 100 percent with up to 1.95 Â 10 6 -fold expansion by using a three-phase culture system that included coculture with a stromal cell line [6]. Later, a protocol was developed to produce red blood cells (RBCs) from human cord blood in the absence of a feeder layer [7]. A different approach, undertaken by Fujimi and co-workers, used a four-phase system which incorporated stromal cells in the first phase and macrophages in the third phase [8]; this yielded more than 10 13 RBCs from one unit of cord blood with a 99.4 percent enucleation efficiency. "
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    • "The most successful of these to date showed enucleation levels of 100 percent with up to 1.95 Â 10 6 -fold expansion by using a three-phase culture system that included coculture with a stromal cell line [6]. Later, a protocol was developed to produce red blood cells (RBCs) from human cord blood in the absence of a feeder layer [7]. A different approach, undertaken by Fujimi and co-workers, used a four-phase system which incorporated stromal cells in the first phase and macrophages in the third phase [8]; this yielded more than 10 13 RBCs from one unit of cord blood with a 99.4 percent enucleation efficiency. "
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    ABSTRACT: An understanding of the metabolic profile of cell proliferation and differentiation should support the optimisation of culture conditions for hematopoietic stem and progenitor cell (HSPC) proliferation, differentiation, and maturation into red blood cells. We have evaluated the key metabolic parameters during each phase of HSPC culture for red blood cell production in serum-supplemented (SS) and serum-free (SF) conditions. A simultaneous decrease in growth rate, total protein content, cell size, and the percentage of cells in the S/G2 phase of cell cycle, as well as an increase in the percentage of cells with a CD71(-)/GpA(+) surface marker profile, indicates HSPC differentiation into red blood cells. Compared with proliferating HSPCs, differentiating HSPCs showed significantly lower glucose and glutamine consumption rates, lactate and ammonia production rates, and amino acid consumption and production rates in both SS and SF conditions. Furthermore, extracellular acidification was associated with late proliferation phase, suggesting a reduced cellular metabolic rate during the transition from proliferation to differentiation. Under both SS and SF conditions, cells demonstrated a high metabolic rate with a mixed metabolism of both glycolysis and oxidative phosphorylation (OXPHOS) in early and late proliferation, an increased dependence on OXPHOS activity during differentiation, and a shift to glycolytic metabolism only during maturation phase. These changes indicate that cell metabolism may have an important impact on the ability of HSPCs to proliferate and differentiate into red blood cells. Copyright © 2015. Published by Elsevier B.V.
    New Biotechnology 05/2015; DOI:10.1016/j.nbt.2015.05.002 · 2.11 Impact Factor
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    • "Due to these challenges, RBC production from hematopoietic stem cells has been a focus in regenerative medicine.6-8 Undifferentiated stem cells confer the advantage of lifelong production in bone marrow. "
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    ABSTRACT: To date, the use of red blood cells (RBCs) produced from stem cells in vitro has not proved practical for routine transfusion. However, the perpetual and widespread shortage of blood products, problems related to transfusion-transmitted infections, and new emerging pathogens elicit an increasing demand for artificial blood. Worldwide efforts to achieve the goal of RBC production through stem cell research have received vast attention; however, problems with large-scale production and cost effectiveness have yet to prove practical usefulness. Some progress has been made, though, as cord blood stem cells and embryonic stem cells have shown an ability to differentiate and proliferate, and induced pluripotent stem cells have been shown to be an unlimited source for RBC production. However, transfusion of stem cell-derived RBCs still presents a number of challenges to overcome. This paper will summarize an up to date account of research and advances in stem cell-derived RBCs, delineate our laboratory protocol in producing RBCs from cord blood, and introduce the technological developments and limitations to current RBC production practices.
    Yonsei medical journal 03/2014; 55(2):304-9. DOI:10.3349/ymj.2014.55.2.304 · 1.26 Impact Factor
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