Expression of Glial and Vimentin Type Intermediate Filaments in Cultures Derived from Human Glial Material

Osborn, Mary; Ludwig-Festl, Monika; Weber, Klaus; Bignami, A.; Dahl, D.; Bayreuther, Klaus

doi:10.1111/j.1432-0436.1981.tb01143.x

Cited by 68 publications

(41 citation statements)

References 30 publications

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“…Table 2 shows discrepant results obtained by immunocytochemical staining of adult human brain cultures. Although minor differences in the percentage of GFAP-positive cells may be attributed to random cell counting errors, it remains unclear why GFAP-positive cells were rare or even absent in adult human brain cultures reported by Osborn et al (1981), Rutka et al (1986) and Perzelova et al (2007). In contrast, Estes et al (1990) and Van der Laan et al (1997) showed a high percentage (more than 80%) of GFAP-positive cells in explanted or dissociated adult human brain cultures.…”

Section: Discussionmentioning

confidence: 98%

GFAP-positive astrocytes are rare or absent in primary adult human brain tissue cultures

et al. 2009

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Traditionally, astrocytes are divided into fibrous and protoplasmic types based on their morphologic appearance. Here the cultures were prepared separately from the adult human cortical gray and white matter of brain biopsies. Both cultures differed only in the number of glial fibrillary acidic protein (GFAP)-positive cells. In the gray matter these were absent or rare, whereas in confluent cultures from the white matter they reached 0.1% of all cells. Three main morphologic types of GFAP-positive cells were found in this study: stellate, bipolar and large flat cells. GFAP-positive cells with two or three long processes mimic a neuron-like morphology. We did not find process-bearing cells expressing neuronal markers (MAP-2, NF, and N-CAM). The conflicting reports concerning GFAP immunostaining and the study dealing with the presence of putative neurons in adult human brain cultures are discussed with respect to these findings. The latter classification of astrocytes into type 1 and type 2 is based on immunostaining to A2B5 antigen: type 1 (GFAP+/A2B5-) and type 2 (GFAP+/A2B5+) astrocytes are proposed to be analogous to protoplasmic and fibrous astrocytes, respectively. In adult human brain cultures we found only small amount of A2B5-positive cells. Double immunofluorescence revealed that astroglial cells of similar fibrous or bipolar shape grown on one coverslip were either GFAP+/A2B5+ or GFAP+/A2B5-. On the other hand, the A2B5+/GFAP-immunophenotype was not observed. These results indicate that in general the cell phenotype from adult human brain tissue is not well established when they are in culture.

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Section: Discussionmentioning

confidence: 98%

GFAP-positive astrocytes are rare or absent in primary adult human brain tissue cultures

et al. 2009

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“…Five (6,7). Coexpression ofvimentin with glial filaments has been reported in cultured astrocytes or glioma cells (8)(9)(10), as well as in astroglia in situ (10)(11)(12). Vimentin can also occur simultaneously with desmin during myogenic differentiation (13,14) and is permanently expressed in cultured baby hamster kidney (BHK-21) cells (15)(16)(17), a line probably derived from an embryonal vascular smooth muscle cell type (18).…”

mentioning

confidence: 99%

Heteropolymer filaments of vimentin and desmin in vascular smooth muscle tissue and cultured baby hamster kidney cells demonstrated by chemical crosslinking.

Quinlan¹,

Franke²

1982

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

147

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Certain smooth muscle cells of blood vessel walls as well as cultured baby hamster kidney cells contain simultaneously two different intermediate-sized filament (IF) proteins, desmin and vimentin. We have examined the question of the occurrence ofboth proteins in the same IF by chemically crosslinking the single cysteine group present in each ofthem. Oxidative crosslinking of filaments present in cytoskeletal preparations with cupric ion complexes of 1,10-phenanthroline resulted in formation ofthree types ofdimers: vimentin-vimentin, desmin-desmin, and vimentin-desmin. These dimers were separated by NaDodSO4/ polyacrylamide gel electrophoresis and characterized by binding of specific antibodies, by one-and two-dimensional gel electrophoresis of monomers obtained after cleavage of the disulfide bond by thiol agents, and by mapping of radioiodinated tryptic peptides. The demonstration ofheterodimers ofvimentin and desmin in vascular smooth muscle tissue of cow and chicken and in baby hamster kidney cells shows that the two proteins can be integrated in the same IF and can be nearest neighbors, oriented with their cysteine residues in a mirror-image symmetry. The possible existence ofheteropolymer IF in other cell types is discussed.tin and desmin has also been reported for Z lines of developing and mature cross-striated muscle, but this is still subject to debate (2)(3)(4)(5)(13)(14)(15).

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“…If not stated otherwise, the studied human cell lines were obtained from gliomas, in most cases probably of astrocytoma origin, including cases of defined glioblastoma multiforme origin (in the terminology and characterizations of the cells and tumors mentioned, we essentially follow the work of Kleihues and Cavenee 2000): U333/MG, U87MG, U138MG, U373MG, T98G (for origins, differentiation markers, and relevant references, see Pontén and Macintyre 1968;Stein 1979;Osborn et al 1981;Achtstätter et al 1986;de Ridder et al 1987;Boda-Heggemann 2005). In addition, we used cultures of cells specifically derived from a glioblastoma multiforme (a kind gift from György Vereb, University of Debrecen, Hungary).…”

Section: Cell Cultures and Cell Culture Linesmentioning

confidence: 99%

“…All are characterized by their differentiation marker complement. Figure 1 presents an example of glioma cells of line U333/ MG, in which all cells contain abundant bundles of intermediate-sized filaments (IFs) of the vimentin type and IFs containing GFAP (for references, see, e.g., Pontén and Westermark 1978;Paetau et al 1980;Osborn et al 1981; for reviews, see Kleihues and Cavenee 2000;Maher et al 2001) and adhaerens junctions based on N-cadherin. Under normal culture conditions, these cells grow in monolayers of flat substratum-adherent cells with typical cytoplasmic actin microfilament cables and focal adhesion attachments fixing these microfilament cables to the basal plasma membrane (Fig.…”

Section: Ve-cadherin In Human Glioma Cell Culturesmentioning

confidence: 99%

Beyond vessels: occurrence and regional clustering of vascular endothelial (VE-)cadherin-containing junctions in non-endothelial cells

2008

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The genes encoding transmembrane glycoproteins of the cadherin family, i.e., the Ca 2+-dependent cell-cell adhesion molecules, are typically expressed in cell-type-or cell-lineage-specific patterns. One of them, vascular endothelial (VE)-cadherin, is widely considered to be specific for vascular endothelia in which it is either the sole or the predominant cadherin, often co-existing with N-cadherin. This specificity of VE-cadherin for vascular endothelial cells is important not only in blood and lymph vessel biology and medicine, but also for cell-type-based diagnoses, notably those of metastatic tumors. Surprisingly, however, we have recently noted the frequent synthesis, surface exposure, and junction assembly of VE-cadherin in certain other cells, in which this glycoprotein is clustered into adherens junctions (AJs), either alone or in combination with N-cadherin and/or cadherin-11. Such cells include mammalian astrocytes and glioma, probably mostly astrocytoma cells growing in culture, and a specific subtype of astrocytoma in situ. Moreover, VE-cadherin synthesis and AJ assembly, plus the regional clustering of such AJs in certain domains, are not clonally fixed but can appear again and again in cells of the progeny of cloned homogeneous-appearing individual cells, thus resulting in clonal cell colonies that are often heterogeneous in their cadherin junction patterns. We discuss the constitutive presence of VE-cadherin in some nonendothelial cells with respect to certain architectural features and possible physiological and pathogenic functions of the cells, and in comparison with recent reports of VE-cadherinpositive melanomas.

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Expression of Glial and Vimentin Type Intermediate Filaments in Cultures Derived from Human Glial Material

Cited by 68 publications

References 30 publications

GFAP-positive astrocytes are rare or absent in primary adult human brain tissue cultures

GFAP-positive astrocytes are rare or absent in primary adult human brain tissue cultures

Heteropolymer filaments of vimentin and desmin in vascular smooth muscle tissue and cultured baby hamster kidney cells demonstrated by chemical crosslinking.

Beyond vessels: occurrence and regional clustering of vascular endothelial (VE-)cadherin-containing junctions in non-endothelial cells

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