DNA synthesis by young adult human-derived astrocytes in vitro

Grenier, Yannick; Ruijs, Theodora C.J.; Olivier, André; Robitaille, Yves; Antel, Jack P.

doi:10.1016/0006-8993(89)91570-9

Cited by 11 publications

(5 citation statements)

References 10 publications

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“…conventional conditions has been limited to a few passages (Grenier et al, 1989;Rutka et al, 1987;Milsted et al, 1990). Several cultures were maintained for 60 to 70 population doublings before proliferation ceased, providing a useful new culture model.…”

Section: Discussionmentioning

confidence: 99%

Serial passage of embryonic human astrocytes in serum‐free, hormone‐supplemented medium

Loo

Sakai

Rawson

et al. 1991

J of Neuroscience Research

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We applied serum-free cell culture methods that allow extended proliferation of mouse astrocyte precursor cells to the multipassage culture of embryonic human brain cells. Cells were cultured in nutrient medium supplemented with insulin, transferrin, epidermal growth factor, fibroblast growth factor, heparin, high-density lipoprotein, and fibronectin. Cultures were maintained for a maximum of 70 population doublings before proliferation ceased. The cells synthesized glial fibrillary acidic protein, an astrocyte marker, and expression of this protein was increased by incubation of the cells with transforming growth factor beta or serum. These results identify extracellular factors important for proliferation and differentiation of embryonic human astrocytes and provide a controlled system for multipassage culture.

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Section: Discussionmentioning

confidence: 99%

Serial passage of embryonic human astrocytes in serum‐free, hormone‐supplemented medium

Loo

Sakai

Rawson

et al. 1991

J of Neuroscience Research

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“…The ghoma cells had a higher rate of proliferation than did the fetal astrocytes; the proliferation of the latter was higher than that of the adult CNS-derived astrocytes . The adult astrocytes can be shown to incorporate significant levels of BrdU in vitro in contrast to the OLs (Grenier et al, 1989).…”

Section: Turget Cell Susceptibility To Tnfa-mediated Injurymentioning

confidence: 99%

Differential susceptibility of human CNS-derived cell populations to TNF-dependent and independent immune-mediated injury

et al. 1995

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We examined whether oligodendrocytes, neurons, and astroglia derived from the human central nervous system differ in susceptibility to injury mediated by tumor necrosis factor (TNF)-alpha and by activated CD4+ T cells acting via a TNF-independent mechanism. Injury was assessed either as cell membrane-directed (lysis), measured by 51chromium (Cr) or lactate dehydrogenase (LDH) release, or nucleus-directed (apoptosis), measured by morphologic features based on propidium iodide (PI) staining and by DNA fragmentation measured by a terminal transferase (TdT)-mediated dUTP biotin nick end labeling technique (TUNEL). TNF did not induce 51Cr or LDH release in any cell targets, but did induce nuclear (66 +/- 2% of cells) and DNA (68 +/- 2% of cells) fragmentation selectively in the oligodendrocytes over 96 hr. At this time, there was no significant loss of oligodendrocyte cell number. Nuclear injury could be induced in neurons by serum deprivation and in malignant astrocytes by the combination of TNF and low serum. CD4+ T cells activated with phytohemagglutin (pha) or anti-CD3 plus interleukin-2 induced significant 51Cr and LDH release in all target cells tested; only pha-activated CD4+ T-cell cocultures showed reduced target cell numbers. Significant nuclear fragmentation was observed only for glioma cells (22 +/- 1% of cells). Differences in susceptibility to different immune effector mechanisms and in the nature of the injury response to the same effector mediator among human CNS-derived neural cells will need to be considered in design of therapeutic strategies aimed at protecting or limiting target cell injury consequent to disease or trauma.

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“…In contrast, Guilian et al (1994) have shown that astrocytes in vitro are involved as supporters of neuronal growth, suggesting that reactive astroglial cells found in glial scar tissue may benefit, and not inhibit, neuronal survival. In other pathologic lesions affecting the human adult CNS, proliferation of astrocytes has been described (Grenier et al, 1989;Norenberg, 1994).…”

Section: Introductionmentioning

confidence: 99%

“…In the past, methods have been developed to isolate viable glial cells from human adult brain tissue. This brain tissue was usually obtained during neurosurgical resection in patients undergoing treatment for tumors or epilepsy (Rutka et al, 1986;Grenier et al, 1989;Estes et al, 1990;Yong et al, 1991;Yong and Antel, 1992), or from organ donors (Whittemore et al, 1993). For the study of many neurological disorders, however, surgical material is not available.…”

Section: Introductionmentioning

confidence: 99%

Establishment of human adult astrocyte cultures derived from postmortem multiple sclerosis and control brain and spinal cord regions: immunophenotypical and functional characterization

Groot¹,

Langeveld

Jongenelen

et al. 1997

J. Neurosci. Res.

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We have successfully established highly enriched astrocyte cultures upon passaging of primary cultures derived from various regions of postmortem human adult brain and spinal cord. Tissues were collected at autopsies with relatively short postmortem times (3-9 hr) from multiple sclerosis (MS) and (normal) control cases. Immunocytochemical analysis showed that primary cultures were composed of colonies of oligoclonal cells that expressed the intermediate filament proteins glial fibrillary acidic protein (GFAP), vimentin, as well as glutamine synthetase (GS). Passaging the astrocytes did not affect their proliferating capacity as monitored by bromodeoxyuridine (BrdU) incorporation. Astrocyte-specific markers were stably expressed for at least 12 passages per individual tissue sample. Large numbers of GFAP-positive astrocytes were obtained from each sample and could be stored frozen and recultured. Very few macrophages/microglial cells (1-3%) were present in the human adult astrocyte cultures, using a panel of macrophage-specific markers. However, the monoclonal antibodies (mAbs KP1, EBM1, 25F9) and lysozyme antiserum directed against lysosomal antigens strongly immunostained cultured astrocytes derived from MS and control cases, implicating that expression of these lysosomal antigens is not restricted to macrophages/ microglial cells in human glial cell cultures. Interestingly, astrocytes derived from active demyelinated MS lesions showed an increased proliferating capacity compared to astrocytes derived from non-lesioned and normal brain and spinal cord regions, as shown with a microculture tetrazolium assay (MTT assay).

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DNA synthesis by young adult human-derived astrocytes in vitro

Cited by 11 publications

References 10 publications

Serial passage of embryonic human astrocytes in serum‐free, hormone‐supplemented medium

Serial passage of embryonic human astrocytes in serum‐free, hormone‐supplemented medium

Differential susceptibility of human CNS-derived cell populations to TNF-dependent and independent immune-mediated injury

Establishment of human adult astrocyte cultures derived from postmortem multiple sclerosis and control brain and spinal cord regions: immunophenotypical and functional characterization

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