Cellular composition of human glial cultures from adult biopsy brain tissue

Gibbons, Hannah M.; Hughes, Stephanie M.; Roon-Mom, Willeke van; Greenwood, J. M.; Narayan, Pritika; Teoh, H. Heng; Bergin, Peter M.; Mee, Edward; Wood, Phil C.; Faull, Richard L.M.; Dragunow, Mike

doi:10.1016/j.jneumeth.2007.07.005

Cited by 51 publications

(65 citation statements)

References 30 publications

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“…The adult human CNS-derived microglia used in this study have biological properties distinct from murine microglia, including lack of proliferation both in vivo and in vitro, and capacity to survive in vitro in the absence of specific growth factor supplementation (27,28). In rodents, TGF-b supplementation in vitro sustains the basal or homeostatic status of these cells, and deletion of TGF-b results in failure of microglia development and survival (16).…”

Section: Discussionmentioning

confidence: 99%

MerTK Is a Functional Regulator of Myelin Phagocytosis by Human Myeloid Cells

Healy

Perron

Won

et al. 2016

The Journal of Immunology

141

160

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Multifocal inflammatory lesions featuring destruction of lipid-rich myelin are pathologic hallmarks of multiple sclerosis. Lesion activity is assessed by the extent and composition of myelin uptake by myeloid cells present in such lesions. In the inflamed CNS, myeloid cells are comprised of brain-resident microglia, an endogenous cell population, and monocyte-derived macrophages, which infiltrate from the systemic compartment. Using microglia isolated from the adult human brain, we demonstrate that myelin phagocytosis is dependent on the polarization state of the cells. Myelin ingestion is significantly enhanced in cells exposed to TGF-β compared with resting basal conditions and markedly reduced in classically activated polarized cells. Transcriptional analysis indicated that TGF-β–treated microglia closely resembled M0 cells. The tyrosine kinase phagocytic receptor MerTK was one of the most upregulated among a select number of differentially expressed genes in TGF-β–treated microglia. In contrast, MerTK and its known ligands, growth arrest-specific 6 and Protein S, were downregulated in classically activated cells. MerTK expression and myelin phagocytosis were higher in CNS-derived microglia than observed in monocyte-derived macrophages, both basally and under all tested polarization conditions. Specific MerTK inhibitors reduced myelin phagocytosis and the resultant anti-inflammatory biased cytokine responses for both cell types. Defining and modulating the mechanisms that regulate myelin phagocytosis has the potential to impact lesion and disease evolution in multiple sclerosis. Relevant effects would include enhancing myelin clearance, increasing anti-inflammatory molecule production by myeloid cells, and thereby permitting subsequent tissue repair.

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Section: Discussionmentioning

confidence: 99%

MerTK Is a Functional Regulator of Myelin Phagocytosis by Human Myeloid Cells

Healy

Perron

Won

et al. 2016

The Journal of Immunology

141

160

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“…Such diploid cells have been described before, following the culture of normal or neoplastic brain tissues from human adults. However, little attention was paid to these cells, which were considered to be glial-like cells or contaminating mesenchymal cells from the meninges or vascular connective tissue [21][22][23].…”

Section: Phenotypic Analysis Of Gascsmentioning

confidence: 99%

Isolation of a new cell population in the glioblastoma microenvironment

et al. 2011

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Glioblastoma (GB) is a highly infiltrative tumor recurring in 90% of cases within a few centimeters of the resection cavity, even in cases of complete tumor resection and adjuvant chemo/radiotherapy. This observation highlights the importance of understanding this special zone of brain tissue surrounding the tumor. It is becoming clear that the nonneoplastic stromal compartment of most solid cancers plays an active role in tumor proliferation, invasion, and metastasis. Very little information, other than that concerning angiogenesis and immune cells, has been collected for stromal cells from GB. As part of a translational research program, we have isolated a new stromal cell population surrounding GB by computer-guided stereotaxic biopsies and primary culture. We named these cells GB-associated stromal cells (GASCs). GASCs are diploid, do not display the genomic alterations typical of GB cells, and have phenotypic and functional properties in common with the cancer-associated fibroblasts (CAFs) described in the stroma of carcinomas. In particular, GASCs express markers associated with CAFs such as fibroblast surface protein, alpha-smooth muscle actin (α-SMA), and platelet-derived growth factor receptor-beta (PDGFRβ). Furthermore, GASCs have a molecular expression profile different from that of control stromal cells derived from non-GB peripheral brain tissues. GASCs were also found to have tumor-promoting effects on glioma cells in vitro and in vivo. The isolation of GASCs in a tumor of neuroepithelial origin was unexpected, and further studies are required to determine their potential as a target for antiglioma treatment.

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“…In order to emphasize a relative increase in the contribution of the assembly-compromised splice-variant GFAPd to total GFAP levels, we chose to display the GFAPd:GFAPa ratio throughout the manuscript. Next, we explored whether U343 astrocytoma cells represent a suitable cell model to further investigate the effect of histone acetylation on GFAP expression, because human primary astrocytes are of limited availability and are prone to loss of their GFAP expression at higher passages (Gibbons et al, 2007). HDAC inhibition with TSA significantly decreased the expression of GFAPa and GFAPd in U343 cells.…”

Section: Histone Acetylation In Astrocytes Reduces Gfap Expression Anmentioning

confidence: 99%

Histone acetylation in astrocytes suppresses GFAP and stimulates a re-organization of the intermediate filament network

Kanski

Sneeboer

Bodegraven

et al. 2014

Journal of Cell Science

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Glial fibrillary acidic protein (GFAP) is the main intermediate filament in astrocytes and is regulated by epigenetic mechanisms during development. We demonstrate that histone acetylation also controls GFAP expression in mature astrocytes. Inhibition of histone deacetylases (HDACs) with trichostatin A or sodium butyrate reduced GFAP expression in primary human astrocytes and astrocytoma cells. Because splicing occurs co-transcriptionally, we investigated whether histone acetylation changes the ratio between the canonical isoform GFAPa and the alternative GFAPd splice variant. We observed that decreased transcription of GFAP enhanced alternative isoform expression, as HDAC inhibition increased the GFAPd:GFAPa ratio. Expression of GFAPd was dependent on the presence and binding of splicing factors of the SR protein family. Inhibition of HDAC activity also resulted in aggregation of the GFAP network, reminiscent of our previous findings of a GFAPd-induced network collapse. Taken together, our data demonstrate that HDAC inhibition results in changes in transcription, splicing and organization of GFAP. These data imply that a tight regulation of histone acetylation in astrocytes is essential, because dysregulation of gene expression causes the aggregation of GFAP, a hallmark of human diseases like Alexander's disease.

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Cellular composition of human glial cultures from adult biopsy brain tissue

Cited by 51 publications

References 30 publications

MerTK Is a Functional Regulator of Myelin Phagocytosis by Human Myeloid Cells

MerTK Is a Functional Regulator of Myelin Phagocytosis by Human Myeloid Cells

Isolation of a new cell population in the glioblastoma microenvironment

Histone acetylation in astrocytes suppresses GFAP and stimulates a re-organization of the intermediate filament network

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