Physiological phenotyping of mammalian cell lines by enzymatic activity fingerprinting of key carbohydrate metabolic enzymes: a pilot and feasibility study
Abstract 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 t...
Main Authors: | , , , , |
---|---|
Format: | Article |
Language: | English |
Published: |
BMC
2019-10-01
|
Series: | BMC Research Notes |
Subjects: | |
Online Access: | http://link.springer.com/article/10.1186/s13104-019-4697-y |
id |
doaj-77f8fee959de4f5f849e4f460251c342 |
---|---|
record_format |
Article |
spelling |
doaj-77f8fee959de4f5f849e4f460251c3422020-11-25T03:44:23ZengBMCBMC Research Notes1756-05002019-10-011211610.1186/s13104-019-4697-yPhysiological phenotyping of mammalian cell lines by enzymatic activity fingerprinting of key carbohydrate metabolic enzymes: a pilot and feasibility studyJulian Geiger0Rebecca Doelker1Sofia Salö2Thomas Roitsch3Louise T. Dalgaard4Department of Science and Environment, Roskilde UniversitySection for Crop Sciences, Department of Plant and Environmental Sciences, Copenhagen Plant Science Centre, University of CopenhagenDepartment of Science and Environment, Roskilde UniversitySection for Crop Sciences, Department of Plant and Environmental Sciences, Copenhagen Plant Science Centre, University of CopenhagenDepartment of Science and Environment, Roskilde UniversityAbstract 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.http://link.springer.com/article/10.1186/s13104-019-4697-yINS-1EEnzyme assays96 well formatAldolaseHexokinaseGlucose-6-phosphate dehydrogenase |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Julian Geiger Rebecca Doelker Sofia Salö Thomas Roitsch Louise T. Dalgaard |
spellingShingle |
Julian Geiger Rebecca Doelker Sofia Salö Thomas Roitsch Louise T. Dalgaard Physiological phenotyping of mammalian cell lines by enzymatic activity fingerprinting of key carbohydrate metabolic enzymes: a pilot and feasibility study BMC Research Notes INS-1E Enzyme assays 96 well format Aldolase Hexokinase Glucose-6-phosphate dehydrogenase |
author_facet |
Julian Geiger Rebecca Doelker Sofia Salö Thomas Roitsch Louise T. Dalgaard |
author_sort |
Julian Geiger |
title |
Physiological phenotyping of mammalian cell lines by enzymatic activity fingerprinting of key carbohydrate metabolic enzymes: a pilot and feasibility study |
title_short |
Physiological phenotyping of mammalian cell lines by enzymatic activity fingerprinting of key carbohydrate metabolic enzymes: a pilot and feasibility study |
title_full |
Physiological phenotyping of mammalian cell lines by enzymatic activity fingerprinting of key carbohydrate metabolic enzymes: a pilot and feasibility study |
title_fullStr |
Physiological phenotyping of mammalian cell lines by enzymatic activity fingerprinting of key carbohydrate metabolic enzymes: a pilot and feasibility study |
title_full_unstemmed |
Physiological phenotyping of mammalian cell lines by enzymatic activity fingerprinting of key carbohydrate metabolic enzymes: a pilot and feasibility study |
title_sort |
physiological phenotyping of mammalian cell lines by enzymatic activity fingerprinting of key carbohydrate metabolic enzymes: a pilot and feasibility study |
publisher |
BMC |
series |
BMC Research Notes |
issn |
1756-0500 |
publishDate |
2019-10-01 |
description |
Abstract 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. |
topic |
INS-1E Enzyme assays 96 well format Aldolase Hexokinase Glucose-6-phosphate dehydrogenase |
url |
http://link.springer.com/article/10.1186/s13104-019-4697-y |
work_keys_str_mv |
AT juliangeiger physiologicalphenotypingofmammaliancelllinesbyenzymaticactivityfingerprintingofkeycarbohydratemetabolicenzymesapilotandfeasibilitystudy AT rebeccadoelker physiologicalphenotypingofmammaliancelllinesbyenzymaticactivityfingerprintingofkeycarbohydratemetabolicenzymesapilotandfeasibilitystudy AT sofiasalo physiologicalphenotypingofmammaliancelllinesbyenzymaticactivityfingerprintingofkeycarbohydratemetabolicenzymesapilotandfeasibilitystudy AT thomasroitsch physiologicalphenotypingofmammaliancelllinesbyenzymaticactivityfingerprintingofkeycarbohydratemetabolicenzymesapilotandfeasibilitystudy AT louisetdalgaard physiologicalphenotypingofmammaliancelllinesbyenzymaticactivityfingerprintingofkeycarbohydratemetabolicenzymesapilotandfeasibilitystudy |
_version_ |
1724515390115020800 |