Novel Method for Efficient Production of Multipotential Hematopoietic Progenitors from Human Embryonic Stem Cells

Ma, Feng; Wang, Dan; Hanada, Shigeo; Ebihara, Yasuhiro; Kawasaki, Hirohide; Zaike, Yuji; Heike, Toshio; Nakahata, Tatsutoshi; Tsuji, Kohichiro

doi:10.1532/ijh97.06203

Cited by 35 publications

(29 citation statements)

References 26 publications

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“…1C). Genes for initial development of endothelial/ hematopoietic cells were expressed early in the coculture, as reported in our previous work (8). The expression of ␤-globin could be detected by day 10, consistent with the immunostaining results.…”

Section: Generation Of Erythroid Cells From Hescs By Coculture With Msupporting

confidence: 90%

“…In a previous study (8), we found that the production of erythroid progenitors from hESCs was greatly enhanced by coculture with mFLSCs. This experimental paradigm provides the opportunity to conduct large-scale investigations of hESC-derived erythropoiesis.…”

Section: Generation Of Erythroid Cells From Hescs By Coculture With Mmentioning

confidence: 80%

“…In our previous report (8), hESC/mFLSC cocultures generated a large number of human hematopoietic progenitors, particularly erythroid and multipotential cells. In the current study, using these clonal erythroid cells, we demonstrated that most hESC-derived E colonies, E bursts, and Mix colonies contained adult-type ␤-globinexpressing erythroid cells, and the percentages of ␤-globin- Progressive maturation of hESC/mFLSC coculture-derived clonal erythroid cells.…”

Section: Discussionmentioning

confidence: 94%

“…Establishment of Murine FL-Derived Stromal Cells. mFLSCs were prepared by as described (8,10). Briefly, FLs were removed from embryonic day 15 (E15) BCL/ Black 6 mice.…”

Section: Methodsmentioning

confidence: 99%

“…The in vitro generation of hematopoietic cells from hESCs has been reported by several studies (4)(5)(6)(7). We also reported a method of hESC coculture with mouse FL-derived stromal cells (mFLSCs) that generated a large quantity of hematopoietic progenitors that could give rise to erythroid cells, providing a means to characterize hESC-derived erythropoiesis (8).…”

mentioning

confidence: 92%

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Generation of functional erythrocytes from human embryonic stem cell-derived definitive hematopoiesis

Ebihara²,

Umeda³

et al. 2008

Proc. Natl. Acad. Sci. U.S.A.

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172

152

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Section: Generation Of Erythroid Cells From Hescs By Coculture With Msupporting

confidence: 90%

Section: Generation Of Erythroid Cells From Hescs By Coculture With Mmentioning

confidence: 80%

Section: Discussionmentioning

confidence: 94%

“…Establishment of Murine FL-Derived Stromal Cells. mFLSCs were prepared by as described (8,10). Briefly, FLs were removed from embryonic day 15 (E15) BCL/ Black 6 mice.…”

Section: Methodsmentioning

confidence: 99%

mentioning

confidence: 92%

See 3 more Smart Citations

Generation of functional erythrocytes from human embryonic stem cell-derived definitive hematopoiesis

Ebihara²,

Umeda³

et al. 2008

Proc. Natl. Acad. Sci. U.S.A.

Self Cite

172

152

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Differentiation of Mast Cells from Embryonic Stem Cells

Kovářová

Koller

2012

CP in Immunology

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In this unit, we describe a simple coculture‐free method for obtaining mast cells from mouse and human embryonic stem (ES) cells. Much of our knowledge regarding the mechanisms by which mast cells are activated comes from studies of mouse bone marrow–derived mast cells. Studies of human mast cells have been hampered by the limited sources from which they can be cultured, the difficulty in introducing specific genetic changes into these cells, and differences between established cultures that reflect the unique genetic makeup of the tissue donor. Derivation of mast cells from embryonic stem cells addresses these limitations. ES‐derived mast cells can be generated in numbers sufficient for studies of the pathways involved in mast cell effector functions. These ES cell–derived mast cells respond to antigens and other stimuli by releasing histamine, cytokines, lipids, and other bioactive mediators. The derivation of human mast cells from ES cells carrying mutations introduced by homologous recombination should provide a novel means of testing the function of genes in both the development and the effector functions of mast cells. Curr. Protoc. Immunol. 97:22F.10.1‐22F.10.16. © 2012 by John Wiley & Sons, Inc.

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Highly efficient and feeder‐free production of subculturable vascular endothelial cells from primate embryonic stem cells

Saeki

Yogiashi

Nakahara

et al. 2008

Journal Cellular Physiology

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The vascular endothelial cell (VEC) differentiation from primate embryonic stem (ES) cells has critical problems: low differentiation efficiencies (<2%) and/or subculture incapability. We report a novel feeder-free culture method for high efficiency production of subculturable VECs from cynomolgus monkey ES cells. Spheres, which were generated from ES cells in the presence of cytokine cocktail, were cultured on gelatin-coated plates. Cobblestone-shaped cells spread out after a few days, which were followed by an emergence of a sac-like structure containing hematopoietic cells. All adherent cells including sac walls cells and surrounding cobblestone cells expressed vascular endothelial cadherin (VE-cadherin) at intercellular junctions. Subculture of these cells resulted in a generation of homogeneous spindle-shaped population bearing cord-forming activities and a uniform acetylated low density lipoprotein-uptaking capacity with von Willbrand factor and endothelial nitric oxide synthetase expressions. They were freeze-thaw-tolerable and subculturable up to eight passages. Co-existence of pericytes or immature ES cells was ruled out. When introduced in a collagen sponge plug implanted intraperitoneally in mice, ES-derived cells recruited into neovascularity. Although percentages of surface VE-cadherin-positive population varied from 20% to 80% as assessed by flow cytometry, the surface VE-cadherin-negative population showed intracellular VE-cadherin expression and mature functions, as we call it as atypical VECs. When sorted, the surface VE-cadherin-positive population expanded as almost pure (>90%) VE-cadherin/PECAM-1-positive VECs by 160-fold after five passages. Thus, our system provides pure production of functional, subculturable and freeze-thaw-tolerable VECs, including atypical VECs, from primate ES cells.

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Novel Method for Efficient Production of Multipotential Hematopoietic Progenitors from Human Embryonic Stem Cells

Cited by 35 publications

References 26 publications

Generation of functional erythrocytes from human embryonic stem cell-derived definitive hematopoiesis

Generation of functional erythrocytes from human embryonic stem cell-derived definitive hematopoiesis

Differentiation of Mast Cells from Embryonic Stem Cells

Highly efficient and feeder‐free production of subculturable vascular endothelial cells from primate embryonic stem cells

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