|
| Status |
Public on May 26, 2016 |
| Title |
mi_static_6_cells |
| Sample type |
RNA |
| |
|
| Source name |
mi_static_6_cells
|
| Organism |
Homo sapiens |
| Characteristics |
cell line: CaCo2
platform_id: Transwell
|
| Treatment protocol |
Transwell inserts with CaCo2 monolayers were either kept on the microplate or put into microfluidic chip. The culture medium microcirculation parameters were ±20 kPa and 2 Hz resulting in the pulsatile (0 - 0.85 µl/s) medium flow with mean flow rate of 4.13 μl/min.
|
| Growth protocol |
The cells were incubated under conditions for differentiation for 7 days in MEM with 10% FBS, 0.1 mM non-essential aminoacids, 0.1% penicillin-streptomycin (all reagents from Gibco, USA) in 5% CO2, 37° C, changed three times.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Qiazol extraction of total RNA was performed according to the manufacturer's instructions.
|
| Label |
biotin
|
| Label protocol |
The procedures for cDNA synthesis and labelling were carried out according to the Ambion WT Expression Kit (Life Technologies, Darmstadt, Germany) using 500 ng total RNA as starting material. ). For miRNA profiling the samples were prepared using FlashTag Biotin HSR RNA Labeling Kit.
|
| |
|
| Hybridization protocol |
The samples were hybridized on GeneChip miRNA 4.0 Arrays (Affymetrix) for 16 h at 48°C.
|
| Scan protocol |
Arrays were washed to remove nonspecifically bound nucleic acids and stained on Fluidics Station 450 (Affymetrix) and then scanned on Gene- Chip Scanner 3000 7G system (Affymetrix).
|
| Description |
miRNA expression data from CaCo2 cells after 24h in Transwell
|
| Data processing |
CEL-files were jointly preprocessed in Affymetrix Expression Console (build 1.4.1.46) using RMA-sketch method for Affymetrix HuGene 1.0 ST arrays and RMA+DAGB-Human_only method for Affymetrix miRNA 4.0 arrays.
|
| |
|
| Submission date |
May 25, 2016 |
| Last update date |
May 26, 2016 |
| Contact name |
Maxim Shkurnikov |
| Organization name |
SRC Bioclinicum
|
| Street address |
2 bld. 85, Ugreshkaya st.
|
| City |
Moscow |
| ZIP/Postal code |
125315 |
| Country |
Russia |
| |
|
| Platform ID |
GPL21572 |
| Series (1) |
| GSE81867 |
miRNA-mediated expression switch of cell adhesion genes driven by microcirculation in chip |
|
