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Insulin and IGF-1 m...
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Adamson, LKarolinska Institutet
(författare)
Insulin and IGF-1 mediated inhibition of apoptosis in CHO cells grown in suspension in a protein-free medium
- Artikel/kapitelEngelska2007
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2007-06-01
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SAGE Publications,2007
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LIBRIS-ID:oai:prod.swepub.kib.ki.se:115810032
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http://kipublications.ki.se/Default.aspx?queryparsed=id:115810032URI
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https://doi.org/10.1177/026119290703500301DOI
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Språk:engelska
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Sammanfattning på:engelska
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When Chinese hamster ovary (CHO) cells were grown in suspension and deprived of serum, 40% of them became apoptotic after 72 hours, as determined by flow cytometry analysis of TUNEL-labelled cells. Cell viability, assessed by erythrocin B staining, decreased correspondingly. An increase in the total fraction of cells expressing interleukin converting enzyme (ICE; caspase 1), B-cell lymphoma 2 protein (Bcl-2,) and Bcl-2 associated x protein (Bax) was shown by antibody probing and subsequent flow cytometry. The p53 tumour suppressor gene product level remained low within the cell population. Insulin-like growth factor-1 (IGF-1) inhibited cell death in a concentration-dependent manner, and at 20ng/ml, cell viability was maintained close to 100% and no apoptotic cells were detected. Also, insulin was shown to inhibit cell death — at 1.0μg/ml, cell viability was 95%, whereas 10% of the cells stained for apoptosis. At the highest concentrations of IGF-1 and insulin, the expression of ICE, Bcl-2 and Bax was fully suppressed, whereas the p53 product level increased, despite still being detectable in a minority of cells. Under these conditions, IGF-1 may increase p53 expression to restrain abnormal cell proliferation. It is concluded that special attention should be paid to exposure and culture conditions that induce acquired susceptibility to a toxic insult, during the development and validation of cell-based assays.
Biuppslag (personer, institutioner, konferenser, titlar ...)
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Walum, E
(författare)
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Karolinska Institutet
(creator_code:org_t)
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Ingår i:Alternatives to laboratory animals : ATLA: SAGE Publications35:3, s. 349-3520261-19292632-3559
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