Physiological phenotyping of mammalian cell lines by enzymatic activity fingerprinting of key carbohydrate metabolic enzymes

a pilot and feasibility study

Julian Geiger, Rebecca Doelker, Sofia Salö, Thomas Roitschd, Louise Torp Dalgaard

Publikation: Bidrag til tidsskriftTidsskriftartikelForskningpeer review

Resumé

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.
OriginalsprogEngelsk
TidsskriftBMC Research Notes
Vol/bind12
Udgave nummer682
ISSN1756-0500
DOI
StatusUdgivet - 2019

Citer dette

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author = "Julian Geiger and Rebecca Doelker and Sofia Sal{\"o} and Thomas Roitschd and Dalgaard, {Louise Torp}",
year = "2019",
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volume = "12",
journal = "BMC Research Notes",
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Physiological phenotyping of mammalian cell lines by enzymatic activity fingerprinting of key carbohydrate metabolic enzymes : a pilot and feasibility study. / Geiger, Julian; Doelker, Rebecca; Salö, Sofia; Roitschd, Thomas ; Dalgaard, Louise Torp.

I: BMC Research Notes, Bind 12, Nr. 682, 2019.

Publikation: Bidrag til tidsskriftTidsskriftartikelForskningpeer review

TY - JOUR

T1 - Physiological phenotyping of mammalian cell lines by enzymatic activity fingerprinting of key carbohydrate metabolic enzymes

T2 - a pilot and feasibility study

AU - Geiger, Julian

AU - Doelker, Rebecca

AU - Salö, Sofia

AU - Roitschd, Thomas

AU - Dalgaard, Louise Torp

PY - 2019

Y1 - 2019

N2 - ObjectiveEnzymatic 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).ResultsEnzyme 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.

AB - ObjectiveEnzymatic 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).ResultsEnzyme 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.

U2 - 10.1186/s13104-019-4697-y

DO - 10.1186/s13104-019-4697-y

M3 - Journal article

VL - 12

JO - BMC Research Notes

JF - BMC Research Notes

SN - 1756-0500

IS - 682

ER -