Immortalized human meibomian gland epithelial cells (IHMGECs) were cultured in the presence or absence of 10% fetal bovine serum, according to published protocols.16 (link)–18 (link) After reaching 80 to 90% confluence (~ 5 × 10/6 (link) well), cells were exposed to ethanol vehicle, linolenic acid (ω-3, 10−5 M; Santa Cruz Biotechnology, Dallas, TX), linoleic acid (ω-6, 10−5 M; Sigma-Aldrich, St. Louis, MO) or linolenic and linoleic acids together (0.5×10−5+ 0.5×10−5M), for 5 to 7 days. Azithromycin (AZM, 10 μg/ml; Santa Cruz Biotechnology) was used as a positive control in all experiments, because this antibiotic has well-defined effects on both the proliferation and differentiation of IHMGECs.19 (link)–22 (link) Following treatment, cells were processed for enumerative, histological and biochemical procedures.
Protocol detail
Effects of Fatty Acids on Meibomian Gland Cells
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Antibiotic
Azithromycin
Cells
Epithelial cells
Ethanol
Fetal bovine serum
Human
Linoleic acid
Linoleic acids
Linolenic acid
Meibomian gland
Corresponding Organization :
Other organizations : Massachusetts Eye and Ear Infirmary, Smith-Kettlewell Eye Research Institute, Harvard University
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Variable analysis
independent variables
- Ethanol vehicle
- Linolenic acid (ω-3, 10^-5 M)
- Linoleic acid (ω-6, 10^-5 M)
- Linolenic and linoleic acids together (0.5×10^-5 + 0.5×10^-5 M)
- Not explicitly mentioned
- Presence or absence of 10% fetal bovine serum
- Cell density (80 to 90% confluence, ~ 5 × 10^6 well)
- Positive control, Azithromycin (AZM, 10 μg/ml)
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