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Gene Expression in MCF10A cells through Differentiation on Transwells

ABSTRACT: To further understand the differences occurring in MCF10A cells as they polarize and differentiate in the Transwell® model, we performed gene expression profiling with Affymetrix Human Genome U133 Plus 2.0 Arrays. Four experimental time points, were sampled: conventional cultures of MCF10A cells grown on plastic (Monolayer) and MCF10A cells plated on Transwells® sampled at three TEER values, 200-300 Ω cm2 (Base), 1400-1600 Ω cm2 (Midpoint), and 3000-3200 Ω cm2 (Plateau). Keywords: Mammary Epithelial Cell Differentiation

ORGANISM(S): Homo sapiens

PROVIDER: GSE10070 | GEO | 2008/01/31

SECONDARY ACCESSION(S): PRJNA108175

REPOSITORIES: GEO

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Gene Expression in MCF10A cells through Differentiation on Transwells

Project description:To further understand the differences occurring in MCF10A cells as they polarize and differentiate in the TranswellÂ® model, we performed gene expression profiling with Affymetrix Human Genome U133 Plus 2.0 Arrays. Four experimental time points, were sampled: conventional cultures of MCF10A cells grown on plastic (Monolayer) and MCF10A cells plated on TranswellsÂ® sampled at three TEER values, 200-300 Î© cm2 (Base), 1400-1600 Î© cm2 (Midpoint), and 3000-3200 Î© cm2 (Plateau). Experiment Overall Design: Cells are grown in monolayer to 90-95% confluency, trypsinized and counted for seeding onto permeable supports (TranswellÂ®, 0.4 Âµm pores, polyester) in normal growth medium. MCF10A cells are seeded on 12-well TranswellsÂ® (Corning) at 105 cells/cm2. Both chambers of media were changed strictly on a 24-hour schedule. Transepithelial electrical resistance (TEER) is measured daily with Epithelial Volt-Ohm Meter (EVOM; World Precision Instruments), prior to media change. Total RNA was isolated at the indicated TEER listed above. Canonical monolayer MCF10A cells served as a control comparison. Each time point was performed on triplicate arrays except for Plateau (n=4).

2008-06-11 | E-GEOD-10070 | biostudies-arrayexpress

Homo sapiens

Project description:Gene Expression in MCF10A cells through Differentiation on Transwells

| PRJNA108175 | ENA

E-GEOD-10070

Project description:Gene Expression in MCF10A cells through Differentiation on Transwells

2014-07-03 | E-GEOD-10070 | ExpressionAtlas

Project description:P8 400 1600

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| PRJNA1099129 | ENA

Cell-matrix adhesion contributes to permeability control in human colon organoids

Project description:Background and aims: Cell-cell adhesion structures (desmosomes and, especially, tight junctions) are known to play important roles in control of transepithelial permeability in the colon. The involvement of cell-matrix interactions in permeability control is less clear. The goals of the present study were to: i) determine if disruption of colon epithelial cell interactions with the extracellular matrix alters permeability control and ii) determine if increasing the elaboration of protein components of cell-matrix adhesion complexes improves permeability control and mitigates the effects of cell-matrix disruption. Results: Treatment of colon organoids with Aquamin® increased the expression of multiple basement membrane and hemidesmosomal proteins as well as keratin 8 and 18. TEER values were higher in the presence of Aquamin® than they were under control conditions. Conclusions: These findings provide evidence that cell-matrix interactions contribute to permeability control in the colon. They suggest that the elaboration of proteins important to cell-matrix interactions can be increased in human colon organoids by exposure to a multi-mineral natural product. Increasing the elaboration of such proteins may help to mitigate the consequences of disrupting cell-matrix interactions on permeability control.

2022-03-14 | PXD026923 | Pride

Regulatory T Cell-Microglia Crosstalk Promotes White Matter Repair after Ischemic Stroke through Osteopontin [microglia]

Project description:We cocultured Tregs in transwells with brain slices collected 5d after tMCAO in the lower compartment for 24h to activate Tregs. Tregs in the transwells were transferred to new wells with primary cultured microglia, and cocultured for 2d. Microglia were then collected for bulk RNAseq analyses. The results revealed roles of activated Tregs in polarizing microglia toward an anti-inflammatory and reparative phenotype.

2021-05-19 | GSE171102 | GEO

Transcriptional response of human enteroids/colonoids on transwells to infection

Project description:The goal of this study was to evaluate the transcriptional response of human enteroids/colonoids on transwells to infections (bacterial and rotavirus). Enteroids/colonoids lines C103, C109, D103, D109, I103, I109, J2 and J11 were plated on transwells coated with Matrigel, differentiated, and inoculated (rotavirus (Ito), bacteria or mock) for 6 or 24 hours. Subsequently, total RNA was isolated and paired-end sequencing was performed.

2021-10-01 | GSE183222 | GEO

Transcriptomics of mouse ileal stem cell-derived epithelium grown on transwells

Project description:Purpose: The goal of this study was to identify transcriptional changes that occur in mouse ileal stem cell-derived epithelium plated on transwells after the establishment of an air-liquid interface (ALI). Methods: Mouse ileal stem cell spheroids were plated on transwells and grown in L-WRN conditioned medium (top and bottom compartment) for 7 days, at which point the top medium was removed to establish ALI. Samples were collected for sequencing on the day of ALI initiation (non-ALI Day 0) and three days post top medium removal (ALI Day 3). Control samples were also collected from transwells submerged for all 10 days of the experiment (non-ALI Day 3) and from the initial stem cell spheroid cultures. Results: Non-ALI Day 0 and non-ALI Day 3 samples had similar transcriptional profiles, whereas ALI transwells showed upregulation of transcriptional factors that induce secretory cell lineages, as well as a general upregulation of metabolic genes involved in oxidative phosphorylation and downregulation of those involved in glycolysis. Conclusions: Removal of L-WRN conditioned medium from the top compartment of transwells plated with stem cell-derived intestinal epithelial cells establishes an air-liquid interface that changes the metabolic profile of the cultures and initiates a differentiation cascade for intestinal secretory cells such as goblet cells and paneth cells.

2019-07-29 | GSE124815 | GEO

Methylation Diet and Lifestyle

Project description:Epigenome analysis of 18 subjects and 20 controls, before, midpoint, and after a change in diet, exercise, and other lifestyle modifications

2023-04-26 | GSE149747 | GEO

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