Physiological phenotyping of mammalian cell lines by enzymatic activity fingerprinting of key carbohydrate metabolic enzymes: a pilot and feasibility study
Jazyk angličtina Země Anglie, Velká Británie Médium electronic
Typ dokumentu časopisecké články
Grantová podpora
FSS 1331-00033B
Danish Medical Research Council
LO1415
Ministry of Education, Youth and Sports of the Czech Republic: National Sustainability Program I
PubMed
31640766
PubMed Central
PMC6805439
DOI
10.1186/s13104-019-4697-y
PII: 10.1186/s13104-019-4697-y
Knihovny.cz E-zdroje
- Klíčová slova
- 96 well format, Aldolase, Enzyme assays, Glucose-6-phosphate dehydrogenase, Hexokinase, INS-1E, Phosphofructokinase, Phosphoglucoisomerase, Phosphoglucomutase,
- MeSH
- aldolasa metabolismus MeSH
- buněčné linie MeSH
- buňky Hep G2 MeSH
- Caco-2 buňky MeSH
- enzymatické testy metody MeSH
- fenotyp MeSH
- fosfofruktokinasy metabolismus MeSH
- fosfoglukomutasa metabolismus MeSH
- fosfotransferasy s alkoholovou skupinou jako akceptorem metabolismus MeSH
- glukosa-6-fosfátdehydrogenasa metabolismus MeSH
- glukosa metabolismus MeSH
- glykolýza * MeSH
- HEK293 buňky MeSH
- hexokinasa metabolismus MeSH
- lidé MeSH
- metabolismus sacharidů * MeSH
- myši MeSH
- nádorové buněčné linie MeSH
- pilotní projekty MeSH
- studie proveditelnosti MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- aldolasa MeSH
- fosfofruktokinasy MeSH
- fosfoglukomutasa MeSH
- fosfotransferasy s alkoholovou skupinou jako akceptorem MeSH
- glukosa-6-fosfátdehydrogenasa MeSH
- glukosa MeSH
- hexokinasa MeSH
- phosphoglucokinase MeSH Prohlížeč
OBJECTIVE: Enzymatic fingerprinting of key enzymes of glucose metabolism is a valuable analysis tool in cell physiological phenotyping of plant samples. Yet, a similar approach for mammalian cell line samples is missing. In this study, we applied semi-high throughput enzyme activity assays that were originally designed for plant samples and tested their feasibility in extracts of six frequently used mammalian cell lines (Caco2, HaCaT, C2C12, HEK293, HepG2 and INS-1E). RESULTS: Enzyme activities for aldolase, hexokinase, glucose-6-phosphate dehydrogenase, phosphoglucoisomerase, phosphoglucomutase, phosphofructokinase could be detected in samples of one or more mammalian cell lines. We characterized effects of sample dilution, assay temperature and repeated freeze-thaw cycles causing potential biases. After careful selection of experimental parameters, the presented semi-high throughput methods could be established as useful tool for physiological phenotyping of cultured mammalian cells.
Department of Science and Environment Roskilde University Universitetsvej 1 4000 Roskilde Denmark
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