Transcriptomics

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Transcription profiling by array of SV40-immortalized human corneal epithelial cells


ABSTRACT: Microarray was used to study global gene expression of a cell culture model based on SV40-immortalized human corneal epithelial (iHCE) cells. The gene expression profile of the cell line was compared to the normal human corneal epithelium. Affymetrix HG-U133A GeneChips® were used for microarray experiments and results were validated by performing RT-qPCR for selected genes. iHCE was found to over- and under-express 22 % and 14 % of the annotated genes, respectively. The results of this study suggest that differences between iHCE cells and normal corneal epithelium are substantial and therefore the use of these cells in corneal research should be considered with caution. SV40-immortalized human corneal epithelial (iHCE) cells were cultured on collagen coated permeable supports for one week using standard culture conditions. Subsequently the apical medium was removed and culturing was continued at the air-liquid interface which allows cells to stratify. Trans-epithelial electrical resistance was followed and total RNA was extracted from cultures with resistance > 300 Ω x cm2. RNA was amplified, labeled with biotin, fragmented and hybridized to the HG-U133A GeneChips® (Affymetrix) according to the protocol of the manufacturer. Three separate experiments with samples from three independent culture batches (iHCE1, iHCE2, iHCE3) were performed. The Global gene expression profile of iHCE was compared to previously published human corneal epithelial tissue (GSE5543) data.

ORGANISM(S): Homo sapiens  

SUBMITTER: Helen C Turner  Arto Urtti  Kati-Sisko Vellonen   Dario Greco   J M Wolosin   Petri Auvinen   Timo Tervo   Marika Häkli      

PROVIDER: E-GEOD-22539 | ArrayExpress | 2010-07-29

SECONDARY ACCESSION(S): GSE22539PRJNA128461

REPOSITORIES: GEO, ArrayExpress

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Publications

Gene expression analysis in SV-40 immortalized human corneal epithelial cells cultured with an air-liquid interface.

Greco Dario D   Vellonen Kati-Sisko KS   Turner Helen C HC   Häkli Marika M   Tervo Timo T   Auvinen Petri P   Wolosin J Mario JM   Urtti Arto A  

Molecular Vision 20101015


To compare the global gene expression profile of stratified epithelia generated in vitro using simian virus 40 (SV40) immortalized human corneal epithelial cells with the previously reported gene expression of normal human corneal epithelia.Immortalized cells expanded in submerged culture were grown in an air-liquid interface of liquid permeable collagen-coated filters to foster stratification and differentiation. Stratified epithelia displaying resistances exceeding 300 Ω·cm2 were dissolved in  ...[more]

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