Evaluation of Cell Harvesting Techniques to Optimize Lipidomic Analysis from Human Meibomian Gland Epithelial Cells in Culture.
cholesteryl esters
human meibomian gland epithelial cells
mass spectrometry
meibomian gland dysfunction
Journal
International journal of molecular sciences
ISSN: 1422-0067
Titre abrégé: Int J Mol Sci
Pays: Switzerland
ID NLM: 101092791
Informations de publication
Date de publication:
06 May 2020
06 May 2020
Historique:
received:
15
04
2020
revised:
05
05
2020
accepted:
05
05
2020
entrez:
10
5
2020
pubmed:
10
5
2020
medline:
2
2
2021
Statut:
epublish
Résumé
The lipidomic analysis of immortalized human meibomian gland epithelial cells (HMGECs) has been proposed as a preclinical model to study meibomian gland dysfunction. An in vitro study was conducted to evaluate neutral lipid recovery following three harvesting techniques and to identify candidate lipid biomarkers of HMGECs. HMGECs were cultured in serum-containing media for two days to promote lipid production. Cells were either harvested by 0.25% trypsin-ethylenediaminetetraacetic acid (EDTA), harvested by 10 mM EDTA, or simultaneously harvested and extracted by 2:1 chloroform-methanol (CM). After extraction by a modified Folch technique, the nonpolar phase was processed and infused into a TripleTOF 5600 mass spectrometer (Sciex, Framingham, MA, USA) with electrospray ionization. MS and MS/MS
Identifiants
pubmed: 32384602
pii: ijms21093277
doi: 10.3390/ijms21093277
pmc: PMC7247324
pii:
doi:
Types de publication
Evaluation Study
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Subventions
Organisme : NIH HHS
ID : RR027822-01
Pays : United States
Organisme : American Optometric Foundation
ID : Beta Sigma Kappa Research Fellowship
Organisme : NEI NIH HHS
ID : K23 EY028629
Pays : United States
Organisme : NEI NIH HHS
ID : P30 EY003039
Pays : United States
Organisme : NEI NIH HHS
ID : EY003039
Pays : United States
Organisme : NCRR NIH HHS
ID : S10 RR027822
Pays : United States
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