Unexpected presence of cytokeratins in human “glia-like” cells

Sivakova, I; Perzelova, A; Kubíková, Eliška; Mráz, P

doi:10.4149/bll_2013_056

Cited by 3 publications

(5 citation statements)

References 19 publications

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“…We detected a high percentage of nestin positive cells in both cell lines, and IF protein assessment confirmed intra and inter-heterogeneity of glioblastoma cell lines. The combination of these and our previous studies [25,27] suggest it is instructive to reconsider the cell-type specificity of IF under culture conditions. STR profiling confirmed the uniqueness of both cell lines and proved their authenticity.…”

Section: Discussionmentioning

confidence: 66%

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High level EGFR amplification in a newly established glioblastoma cell line 170-MG-BA

MacLeod

Schneider

Sivakova

et al. 2019

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Glioblastoma multiforme is a highly invasive and incurable primary brain tumor. The most frequent genetic alteration therein is amplification of the epidermal growth factor receptor (EGFR) gene, the target of current clinical trials. However, EGFR amplification is poorly represented in glioblastoma cell lines. From the 30 cultures attempted herein, we were able to establish two glioblastoma permanent cell lines. The remaining cultures showed limited life span and underwent senescence between passage numbers (PN) 8 to 15. Our newly established glioblastoma cell lines, designated 170-MG-BA and 538-MG-BA, both originated between PN 3 and 5 when areas of smaller, more rapidly proliferating cells appeared. Both cell lines showed similar rates of growth, moderate morphological differences, cytoskeletal heterogeneity and multiple chromosome rearrangements. Analysis by molecular cytogenetics and comparative genomic hybridization (aCGH) revealed two copies of a stable marker chromosome in 170-MG-BA cells effecting focal amplification at 7q11 of the EGFR locus. Comparative RqPCR analysis confirmed that EGFR was uniquely highly expressed in 170-MG-BA cells. Combined targeted expression analysis and aCGH data excluded the recurrent EGFRvIII activating mutation. In contrast, EGFR expression in 538-MG-BA cells which lacked genomic EGFR amplification was not raised. Immunofluorescent staining showed high EGFR protein expression only in the 170-MG-BA cells. Cytogenetic, genomic and transcriptional analyses then confirmed high-level genomic amplification and transcriptional upregulation of wild type EGFR in 170-MG-BA; the first conventional cell line model for investigating the biology and targeted therapy of this key alteration in glioblastoma. Both cell lines are freely available from the DSMZ cell repository.

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Section: Discussionmentioning

confidence: 66%

“…Although specific for normal and neoplastic epithelial cells, cytokeratins (CK) are found in glioma tissue [23]. Recently we described a subpopulation of CK-positive cells in previously established glioblastoma cells lines [24] and also in normal human "glial-like" cells [25].…”

Section: Discussionmentioning

confidence: 99%

High level EGFR amplification in a newly established glioblastoma cell line 170-MG-BA

MacLeod

Schneider

Sivakova

et al. 2019

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“…In this study CK-positive cells were present in 25/28 glioblastoma cultures at various percentages. Previously we described the unexpected presence of CK in 8/10 human brain cultures where 0.1 to 70 % of "glia-like" cells positively stained with monoclonal anti-pan CK antibodies (9). Celullar dediferentiation is hallmark of cancer cells, however, CK expression in cultured normal brain cells is a sign of cellular dedifferentiaton which recapitulates the ectodermal origin of brain tissue.…”

Section: Discussionmentioning

confidence: 91%

“…Normal cultures contained only minor populations of morphologically distinct glial cell types (2 to 5 %) expressing specifi c markers accompanying a predominant population of GFAP-/vimentin+ "glia-like" cells (8). Similarly in normal "glia-like" and glioblastoma cultures the cells expressed nestin or cytokeratins at various percentages (8,9). Differences were observed only in percentages of GFAP-positive cells.…”

Section: Discussionmentioning

confidence: 99%

Short-term glioblastoma cultures may contain normal “glia-like“ cells

Sivakova¹,

MacLeod²,

Mráz³

et al. 2019

BLL

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OBJECTIVES: Currently used glioblastoma cultures have many disadvantages and are being replaced by short-term cultures. However, these may include normal brain cells. BACKGROUND: A comparative model of normal and glioma cultures is lacking. A signifi cant contributory factor is because cultures from adult human brain contain small amounts of cells with glial phenotypes. The predominant population of fl at or spindle shaped cells does not express glial markers and are often termed as "glia-like". METHODS: Cryopreserved glioblastoma cultures from 28 bioptic samples were examined by immunofl uorescence using antibodies to intermediate fi laments (IF): glial fi brillary acidic protein (GFAP), cytokeratins (CK), nestin (Nes), vimentin (Vim) and neurofi laments (NF). RESULTS: In short-term glioblastoma cultures GFAP-positive cells occured at higher percentages in 3/28 cultures and in lower percentages in further 5 cultures. Subpopulation of nestin positive cells were observed in all cultures and CK-positive cells were found in 25/28 cultures. All cells in all cultures were positively stained only for vimentin and negatively for NF. Cells grew slowly in 5 cultures which showed early proliferation arrest between passages 7 to 8. A further 23 cultures showed growth arrest by passages 10 to 15. CONCLUSION: The presence of normal cells in short-term glioblastoma cultures may be caused by the infi ltrative growth of these tumors. Our comparative analysis of morphological, growth and cytoskeletal properties revealed similarities between glioblastoma and normal brain cultures. In this study, the majority (28/30) of short-term glioblastoma showed limited life spans, similar to normal cells lacking spontaneous immortalization. The use of short-term glioblastoma cultures has two main problematic areas: cultures may contain a major subpopulation of normal "glia-like" cells; or they may contain the inital phases of spontaneously immortalized glioblastoma cells bearing properties of permanent cell lines (Tab. 1, Fig. 2 , Ref. 19). Text in PDF www.elis.sk.

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“…The pan-cytokeratin (Pan-CK), or cytokeratin, test has become the cornerstone of evaluating evidence of epithelial differentiation, and it is probably the most commonly used IHC test in almost every general or subspecialty practice in pathology and cytology (30). Cytokeratins (CKs) are intermediate filaments (IFs) that are specific to normal and neoplastic epithelial cell differentiation (31). They play an important role as structural and functional proteins of the cytoskeleton.…”

Section: Pan-ckmentioning

confidence: 99%

Low 06-Methylguanine-DNA Methytransferase (MGMT) and Pan-Cytokeratin (PAN-CK) Expression via Immunohistochemistry in Pituitary Adenomas

et al. 2017

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Introduction. Pituitary adenomas (PA) are the third most common intracranial tumors, with an incidence rate of 10-15%. More than half are invasive, infiltrating adjacent structures. The primary objective of this project was to determine whether MGMT expression is associated with the invasiveness of PA. Material and Method. All patients who underwent surgical decompression consecutively between 2007-2012 were included. All data were obtained from the case records. Formalin-fixed paraffin-embedded (FFPE) tissue specimens were stained with hematoxylin and eosin (HE) and then examined via light microscope. Paraffin blocks that lacked necrosis and hemorrhage were chosen for histologic examination. In addition to an immunoprofile battery that consisted of Ki-67 and p53, MGMT, S-100 and Pan-CK were evaluated as well. Results. The subjects included 25 women and 15 men. The mean age was 48.9 ± 14.5 years. Of these, 63% of cases involved the invasion of adjacent structures. Of the PA, 17 (42%) were non-functioning pituitary adenomas (NFPA). There was a statistically significant relationship between the invasiveness and Ki-67, p53, MGMT expression, and prolactinoma. Gonodotropinomas were mostly non-invasive. FPAs presented invasive features more frequently than NFPAs. Pan-CK was positive in GH-secreting adenomas but negative in FSH-and LH-secreting adenomas. Conclusion. Ki-67 and p53 in lower expression level can be used for evaluating invasiveness but not for recurrence. MGMT expression can be a useful IHC indicator for invasiveness. However, Pan-CK cannot be used for invasiveness or aggressiveness.

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Unexpected presence of cytokeratins in human “glia-like” cells

Cited by 3 publications

References 19 publications

High level EGFR amplification in a newly established glioblastoma cell line 170-MG-BA

High level EGFR amplification in a newly established glioblastoma cell line 170-MG-BA

Short-term glioblastoma cultures may contain normal “glia-like“ cells

Low 06-Methylguanine-DNA Methytransferase (MGMT) and Pan-Cytokeratin (PAN-CK) Expression via Immunohistochemistry in Pituitary Adenomas

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