Personal tools

Epithelial to mesenchymal

From FANTOM5_SSTAR

Jump to: navigation, search

Series:IN_VITRO DIFFERENTIATION SERIES
Species:Human (Homo sapiens)
Genomic View:Zenbu
Expression table:FILE
Link to TET:TET
Sample providers :Soichi Ogishima
Germ layer:mesoderm
Primary cells or cell line:cell line
Time span:60 hours
Number of time points:21


CollapseOverview

Epithelial\u2013mesenchymal transition (EMT) is a process of transition from epithelial cells to mesenchymal cells characterized by loss of cell adhesion and polarity, repression of E-cadherin expression, gain of cell motility and nvasive properties. EMT is essential for numerous developmental processes including mesoderm formation and neural tube formation. EMT is also essential for wound healing, organ fibrosis and initiation of metastasis for cancer progression. During the process of epithelial to mesenchymal transition, epithelial cells transforms to mesenchymal cells, and change their shapes. Mesenchymal cells get to gather and form focus. To clarify the transcriptional networks regulating EMT, we obtained time-course sample of human ARPE-19 epithelial to mesenchymal transition. After preculture for 7 days on 10cm dishes and cell seeding and cell culture for 5 days on 6-well glass bottom plate, we induced epithelial to mesenchymal transition on human ARPE-19 cells by TT-mixture of TNF\u03b1 and TGF\u03b22. Triplication for RNA extraction and 1 for imaging. We sampled total RNA of human ARPE-19 cells along with time-course of epithelial to mesenchymal transition.

Sample description

ARPE-19 cells were cultured in Dulbecco\u2019s Modified Eagle\u2019s Medium/Nutrient Mixture F-12 medium (Sigma-Aldrich) with 10% fetal bovine serum in a CO2 incubator at 37 ｰC. After 5 days of preculture, cells were treated with human recombinant TGF-\u03b22 and human recombinant TNF-\u03b1 and continuously incubated in serum free medium at 37 ｰC to induce EMT. At 21 different time points (0 min, 15 min, 30 min, 45 min, 60 min, 80 min, 100 min, 2 hr, 2.5 hr, 3 hr, 3.5 hr, 4 hr, 5 hr, 6 hr, 7 hr, 8 hr, 12 hr, 16 hr, 24 hr, 42 hr and 60 hr), cells were dissolved in lysis reagent and collected. For the samples of 0 min, cells were treated with PBS without TGF-\u03b22 and TNF-\u03b1. Using miRNeasy Mini Kit (QIAGEN), total RNA was extracted from each cell lysate according to a manufacturer\u2019s protocol.

Figure 1: Sample protocol


Figure 2: Cell morphology

Quality control


Figure 3: CAGE expression of marker genes in TPM.

Profiled time course samples

Only samples that passed quality controls (Arner et al. 2015) are shown here. The entire set of samples are downloadable from FANTOM5 human / mouse samples


Retrieved from "http://f5-webapp4.gsc.riken.jp/resource_browser/index.php?title=Epithelial_to_mesenchymal&oldid=4578783"