Computational protocol: Serum-Induced Keratinization Processes in an Immortalized Human Meibomian Gland Epithelial Cell Line

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Protocol publication

[…] Mass spectrometric analysis was performed using a modification of the method of Brown et al. []. Mass spectra were acquired using a chip based nano-electrospray ionization source (TriVersa Nanomate, Advion, Ithaca, NY, USA) coupled to a hybrid linear ion trap-triple quadrupole mass spectrometer (QTRAP 5500, ABSCIEX, Foster City, CA, USA). Data were analyzed with LipidView (ABSCIEX) software version 1.1 as per the methods of Brown et al. and Tran et al. Precursor ion and neutral loss scans covering the most abundant lipid classes found in meibum (wax ester (WE), cholesterol ester (CE)) as well as abundant lipid classes in tissue (phosphatidylethanolamine (PE), phosphatidylserine (PS), phosphatidylcholine (PC), sphingomyelin (SM), ceramide (Cer), free cholesterol (Chol), diacylglycerol (DAG) and triacylglycerol (TAG)) were utilized to give coverage of both meibum and cellular lipids. MS conditions are provided in . [...] Data are expressed as mean ± SEM. All variables were normally distributed according to Kolmogorov–Smirnov test. After evaluating values for normal distribution we performed one-way ANOVA statistics. For the interpretation of the results we used either Bonferroni or Games- Howell post hoc test depending on Levene’s test for homogeneity of variances. All data were analyzed by IBM SPSS Statistics software package version 20.0 (IBM SPSS). A p-value less than 0.05 was considered statistically significant. […]

Pipeline specifications

Software tools LipidView, SPSS
Applications Miscellaneous, MS-based untargeted lipidomics
Organisms Homo sapiens
Chemicals Cholesterol Esters, Glucose, Eicosapentaenoic Acid