Generation of Lymphohematopoietic Cells from Embryonic Stem Cells in Culture

Nakano, Toru; Kodama, Hiroaki; Honjo, Tasuku

doi:10.1126/science.8066449

Cited by 806 publications

(606 citation statements)

References 28 publications

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“…We first examined the supportive capacity of different cell lines and found the OP9 and 3T3-L1 preadipocytic cell lines to be superior. The OP9 cells line is a commonly used cell line to support primitive cells, including those of the hematopoietic and embryonic lineages [39,40]. While this demonstrated the crucial role of cell contact for the augmented support, we further wished to analyze mature adipocytes in a primary setting.…”

Section: Discussionmentioning

confidence: 99%

Adipocytic Cells Augment the Support of Primitive Hematopoietic Cells In Vitro But Have No Effect in the Bone Marrow Niche Under Homeostatic Conditions

Spindler

Tseng

Zhou

et al. 2014

Stem Cells and Development

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Mesenchymal stem cells (MSCs), as well as osteoblastic cells derived from these MSCs, have been shown to be key components of the hematopoietic stem cell (HSC) niche. In this study, we wished to examine whether other cell types that are known to differentiate from MSCs similarly regulate the stem cell niche, namely cells of the adipocyte lineage. Recent studies have examined the role that adipocytes play in the biology of the HSCs in different bone locations and in transplantation settings; however, none have examined their role under homeostatic conditions. We compared the ability of adipocytic and nonadipocytic cell lines to support primitive hematopoietic cells in vitro. Preadipocytic cell lines demonstrated enhanced support of hematopoietic cells. Similarly, primary bone marrow (BM) cells treated with troglitazone, a drug that enhances adipogenesis, also demonstrated augmented support over control-treated stromal cells. We further examined the effects of increased adipocyte number in vivo under homeostatic conditions using troglitazone treatment and found that these alterations had no effect on HSC frequency. Taken together, we demonstrate that cells of the adipocyte lineage promote the ability of stromal cells to support primitive hematopoietic cells in vitro, yet alterations of adipocyte number and volume in vivo have no effect. These data suggest that adipocytes are not a component of the adult BM HSC niche under homeostatic conditions.

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Section: Discussionmentioning

confidence: 99%

Adipocytic Cells Augment the Support of Primitive Hematopoietic Cells In Vitro But Have No Effect in the Bone Marrow Niche Under Homeostatic Conditions

Spindler

Tseng

Zhou

et al. 2014

Stem Cells and Development

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“…1a,b). Differentiation of ES cells to macrophages was started by cocultivation of ES cells on OP9 stroma cells (Nakano et al 1994) as described (Schroeder et al 2003). At day 8 of differentiation, suspension cells were transferred to cell culture flasks using medium containing macrophage colony-stimulating factor (M-CSF) and interleukin 3 (IL-3).…”

Section: Cellsmentioning

confidence: 99%

Three-dimensional positioning of genes in mouse cell nuclei

et al. 2008

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To understand the regulation of the genome, it is necessary to understand its three-dimensional organization in the nucleus. We investigated the positioning of eight gene loci on four different chromosomes, including the β-globin gene, in mouse embryonic stem cells and in in vitro differentiated macrophages by fluorescence in situ hybridization on structurally preserved nuclei, confocal microscopy, and 3D quantitative image analysis. We found that gene loci on the same chromosome can significantly differ from each other and from their chromosome territory in their average radial nuclear position. Radial distribution of a given gene locus can change significantly between cell types, excluding the possibility that positioning is determined solely by the DNA sequence. For the set of investigated gene loci, we found no relationship between radial distribution and local gene density, as it was described for human cell nuclei. We did find, however, correlation with other genomic properties such as GC content and certain repetitive elements such as long terminal repeats or long interspersed nuclear elements. Our results suggest that gene density itself is not a driving force in nuclear positioning. Instead, we propose that other genomic properties play a role in determining nuclear chromatin distribution.

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“…Approximately 5 ϫ 10 4 P-Spderived cells were suspended in 300 L of serum-free StemPro media (Life Technologies, Gaithersburg, MD) supplemented with 50 ng/mL stem-cell factor (SCF), 5 ng/mL interleukin-3 (IL3; gifts from Kirin Brewery, Takasaki, Japan), and 10 ng/mL mouse oncostatin M (R&D Systems, Minneapolis, MN). Single-cell suspensions were seeded on preplated OP9 stromal cells 15 in the 24-well plate, followed by incubation at 37°C in a 5% CO 2 incubator. Images were visualized with a Nikon Eclipse TE2000-U microscope equipped with 40ϫ/0.60 and 10ϫ/0.30 NA objective lenses (Nikon, Tokyo, Japan), and were captured with a C5810 camera (Hamamatsu Photonics, Hamamatsu, Japan).…”

Section: Mice and Embryosmentioning

confidence: 99%