Schepens Eye Research Institute, Boston, Massachusetts, United States.
Proteomics and Mass Spectrometry Facility, University of Massachusetts Medical School, Shrewsbury, Massachusetts, United States.
Invest Ophthalmol Vis Sci. 2014 May 27;55(6):3866-77. doi: 10.1167/iovs.13-13407.
We hypothesize that culturing immortalized human meibomian gland epithelial cells in serum-containing medium will induce their differentiation. The purpose of this investigation was to begin to test our hypothesis, and explore the impact of serum on gene expression and lipid accumulation in human meibomian gland epithelial cells.
Immortalized and primary human meibomian gland epithelial cells were cultured in the presence or absence of serum. Cells were evaluated for lysosome and lipid accumulation, polar and neutral lipid profiles, and gene expression.
Our results support our hypothesis that serum stimulates the differentiation of human meibomian gland epithelial cells. This serum-induced effect is associated with a significant increase in the expression of genes linked to cell differentiation, epithelium development, the endoplasmic reticulum, Golgi apparatus, vesicles, and lysosomes, and a significant decrease in gene activity related to the cell cycle, mitochondria, ribosomes, and translation. These cellular responses are accompanied by an accumulation of lipids within lysosomes, as well as alterations in the fatty acid content of polar and nonpolar lipids. Of particular importance, our results show that the molecular and biochemical changes of immortalized human meibomian gland epithelial cells during differentiation are analogous to those of primary cells.
Overall, our findings indicate that immortalized human meibomian gland epithelial cells may serve as an ideal preclinical model to identify factors that control cellular differentiation in the meibomian gland.
我们假设在含血清的培养基中培养永生化的人眼睫毛腺上皮细胞会诱导其分化。本研究旨在初步验证我们的假设,并探讨血清对人眼睫毛腺上皮细胞基因表达和脂质积累的影响。
在有或没有血清的情况下培养永生化和原代人眼睫毛腺上皮细胞。评估细胞内溶酶体和脂质积累、极性和中性脂质谱以及基因表达。
我们的结果支持我们的假设,即血清刺激人眼睫毛腺上皮细胞的分化。这种血清诱导的作用与与细胞分化、上皮发育、内质网、高尔基体、囊泡和溶酶体相关的基因表达显著增加有关,与细胞周期、线粒体、核糖体和翻译相关的基因活性显著降低有关。这些细胞反应伴随着溶酶体内脂质的积累,以及极性和非极性脂质中脂肪酸含量的改变。特别重要的是,我们的结果表明,在分化过程中永生化人眼睫毛腺上皮细胞的分子和生化变化与原代细胞相似。
总体而言,我们的发现表明,永生化人眼睫毛腺上皮细胞可能成为一种理想的临床前模型,用于鉴定控制眼睫毛腺细胞分化的因素。