Project description:Analysis of gene expression profile changes of immortalized human meibomian gland epithelial cells under differentiation. Results provide important information of the response of human meibomian gland epithelial cells to different differentiation media, such as serum containing medium, serum-free medium, serum-free medium with 1.2mM Ca2+.
Project description:Analysis of gene expression profile changes of immortalized human meibomian gland epithelial cells under differentiation. Results provide important information of the response of human meibomian gland epithelial cells to different differentiation media, such as serum containing medium, serum-free medium, serum-free medium with 1.2mM Ca2+. Primary cultured human meibomian gland epithelial cells and hTERT immortalized human meibomian gland epithelial cells (3 cells/condition/experiment) were grown in keratinocyte serum-free medium (SFM, Invitrogen-Gibco, Grand Island, NY) on 75cm2 to reach confluence, then change to SFM, SFM with 1.2mM Calcium, or serum containing medium (10% fetal bovine serum in an equal volume of DMEM and Ham’s F12 with 10ng/ml EGF) for 14 days. Total RNA was extracted from the above cultures. Gene expression was analyzed using Illumina HumanHT-12 v3 Expression Beadchips (San Diego, CA).
Project description:Analysis of growth factor influence on immortalized human meibomian gland epithelial cells at gene expression level. Growth factors play a critical role in the proliferation and differentiation of sebaceous gland epithelial cells. Given that the meibomian gland is a large sebaceous gland, we hypothesize that growth factors exert analogous effects on human meibomian gland epithelial cells. Results provide important information of the response of human meibomian gland epithelial cells to epidermal growth factor (EGF), bovine pituitary extract (BPE), and both, such as up- or down- regulated genes involved in lipid metabolic process and cell cycle.
Project description:Genome-wide analysis of dihydrotestosterone (DHT) induced changes in gene expression in immortalized human meibomian gland epithelial cells. Analysis of regulation of immortalized human meibomian gland epithelial cells by dihydrotestosterone at gene expression level. The hypothesis tested in the present study was that the androgen-eye interaction in ocular surface epithelial cells like meibomian gland cells is influenced by androgens through regulation of the expression of multiple genes. Results provide important information of the differential regulation of numerous genes in response to dihydrotestosterone incubation of immortalized human meibomian gland epithelial cells. Total RNA was obtained from immortalized human meibomian gland epithelial cells treated for 72 hours with 10 nM dihydrotestosterone (n=3) or vehicle (n=3). The RNA was then used with Illumina HumanHT-12 v3 Expression BeadChips to determine the effect of DHT on gene expression in an immortalized human meibomian epithelial cell line developed in our laboratory.
Project description:Genome-wide analysis of dihydrotestosterone (DHT) induced changes in gene expression in immortalized human meibomian gland epithelial cells. Analysis of regulation of immortalized human meibomian gland epithelial cells by dihydrotestosterone at gene expression level. The hypothesis tested in the present study was that the androgen-eye interaction in ocular surface epithelial cells like meibomian gland cells is influenced by androgens through regulation of the expression of multiple genes. Results provide important information of the differential regulation of numerous genes in response to dihydrotestosterone incubation of immortalized human meibomian gland epithelial cells.
Project description:Analysis of growth factor influence on immortalized human meibomian gland epithelial cells at gene expression level. Growth factors play a critical role in the proliferation and differentiation of sebaceous gland epithelial cells. Given that the meibomian gland is a large sebaceous gland, we hypothesize that growth factors exert analogous effects on human meibomian gland epithelial cells. Results provide important information of the response of human meibomian gland epithelial cells to epidermal growth factor (EGF), bovine pituitary extract (BPE), and both, such as up- or down- regulated genes involved in lipid metabolic process and cell cycle. Human meibomian gland epithelial cells were immortalized in our lab with a retrovirus containing telomerase reverse transcriptase (hTERT). hTERT immortalized cells (3 wells /condition / experiment) were seeded onto 6-well plates at the density of 3.76M-CM-^W104cells /well in SFM basal medium, SFM basal medium with epidermal growth factor (EGF; 5 ng/ml), SFM basal medium with bovine pituitary extract (BPE; 50 M-BM-5g/ml), and SFM basal medium with 5ng/ml EGF plus 50 M-BM-5g/ml BPE (KGM). Total RNA was extracted from cultures in SFM basal medium at day 2, and from cultures in SFM basal medium with BPE, basal medium with EGF, and SFM basal medium with EGF plus BPE at day 7.
Project description:The objective of this study is to identify human meibomian gland genes that may promote the development and/or progression of meibomian gland dysfunction.