Characterization of microglia induced from mouse embryonic stem cells and their migration into the brain parenchyma

Tsuchiya, T.; Park, Kae Chang; Toyonaga, Shinichi; Yamada, Shoko M.; Nakabayashi, Hiromichi; Nakai, Eiichi; Ikawa, Naoki; Furuya, Masato; Tominaga, Akira; Shimizu, Katsuji

doi:10.1016/j.jneuroim.2004.10.025

Cited by 40 publications

(35 citation statements)

References 67 publications

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“…Our study was initiated by a pioneering work from Tsuchiya et al, who suggested that a small population of microglial-like cells can be found next to neural cells differentiated out of mouse EBs (Tsuchiya et al, 2005). Their differentiation protocol was based on a protocol for the generation of dopaminergic neurons (Lee et al, 2000).…”

Section: Discussionmentioning

confidence: 99%

“…Using a protocol of neuronal differentiation, they succeeded in isolation of a subpopulation of cells by a density gradient method that were positively stained for IBa11 and CD451. The isolated cells showed morphological characteristics of primary microglia after transplantation in mice, but were not described to survive and proliferate in culture (Tsuchiya et al, 2005).…”

Section: Introductionmentioning

confidence: 94%

“…Recently, differentiation of microglia-like cells from ES cells was described (Tsuchiya et al, 2005). Using a protocol of neuronal differentiation, they succeeded in isolation of a subpopulation of cells by a density gradient method that were positively stained for IBa11 and CD451.…”

Section: Introductionmentioning

confidence: 99%

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Microglial precursors derived from mouse embryonic stem cells

Napoli

Kierdorf

Neumann

2009

Glia

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Microglia are resident immune cells of the central nervous system. They can be directly isolated from the brain or from mixed postnatal glial cultures. Isolation of primary microglia is inefficient due to low yield. The cell line BV2 was used as a substitute for primary microglia, but BV2 are oncogenically transformed cells. Here, we established a protocol to generate microglial precursor lines from mouse embryonic stem (ES) cells. Microglial precursor cells were obtained from murine ES cells by differentiation of embryoid bodies to microglia within a mixed brain culture. Several independent ES cell-derived microglial precursor (ESdM) lines were generated and characterized by flow cytometry, immunocytochemistry, and functional assays. All ESdM showed expression of IBa1, CD11b, CD45, F4/80, CD49d, and CD29, but were negative for cKit and CD34. Stimulation with interferon-gamma or lipopolysaccharide (LPS) demonstrated upregulation of proinflammatory cytokine gene transcription including nitric oxide synthase-2, interleukin-1beta, and tumor necrosis factor-alpha at levels comparable to primary microglia. The ESdM showed efficient and rapid phagocytosis of microsphere beads, which was increased after stimulation with LPS. ESdM expressed the chemokine receptor CX3CR1 and demonstrated directed migration toward the ligand CX3CL1. After in vivo transplantation into postnatal brain tissue, ESdM showed engraftment as cells with a microglial phenotype and morphology. Thus, ESdM are stable proliferating cells substantially having most characteristics of primary microglia and therefore being a suitable tool to study microglial function in vitro and in vivo.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 94%

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Microglial precursors derived from mouse embryonic stem cells

Napoli

Kierdorf

Neumann

2009

Glia

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“…To this end, some groups have directly tested whether in vitro differentiated microglia can home to the CNS following intravenous injection. 31 The injected cells were indeed found to preferentially home to the CNS (primarily the hippocampus and corpus callosum); however, these studies did not determine how long these cells remained in the CNS, nor whether they fully integrated into the CNS parenchyma. 31 These studies demonstrate the potential of a small number of differentiated microglia to traffic across an intact blood-brain barrier into the CNS, but do not address whether the bone marrow routinely generates cells able to preferentially home to the CNS and then differentiate into microglia.…”

Section: A Rose By Any Other Name…?mentioning

confidence: 94%

A Rose by Any Other Name? The Potential Consequences of Microglial Heterogeneity During CNS Health and Disease

et al. 2007

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SummaryMicroglial activation and macrophage infiltration into the CNS are common features of CNS autoimmune disease and of chronic neurodegenerative diseases. Because these cells largely express an overlapping set of common macrophage markers, it has been difficult to separate their respective contributions to disease onset and progression. This problem is further confounded by the many types of macrophages that have been termed microglia. Several approaches, ranging from molecular profiling of isolated cells to the generation of irradiation chimeric rodent models, are now beginning to generate rudimentary definitions distinguishing the various types of microglia and macrophages found within the CNS and the potential roles that these cells may play in health and disease.

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“…Recently, a protocol for the generation of microglia derived from mES cells was developed (Beutner et al, 2010;). This effort was initiated by a work from Tsuchiya and colleagues who succeeded in the differentiation of microglial-like cells derived from mES cells (Tsuchiya et al, 2005). During their neuronal differentiation based on a protocol for the generation of dopaminergic neurons (Lee et al, 2000) they found a population positive for Iba1 and CD45.…”

Section: Embryonic and Induced Pluripotent Stem Cell-derived Microgliamentioning

confidence: 99%