Dataset features


Application: RNA-seq analysis, miRNA array analysis
Number of samples: 12
Release date: May 26 2016
Last update date: Jul 26 2018
Access: Public
Diseases: Colonic Neoplasms
Dataset link miRNA-mediated expression switch of cell adhesion genes driven by microcirculation in chip

Experimental Protocol

Undifferentiated Caco2 cells were seeded on individual polyether membrane inserts with 0.143 cm2 surface area and 1.0 μm pore diameter, cut out from HTS Transwell®-96 well permeable support (Corning Inc., USA), with cell density approximately 60,000 cells/well. 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 in 5% CO2, 37° C, changed three times. 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. After 24-h incubation the cells were lysed in 700 μl of Qiazol lysis reagent (Qiagen, Germany) and subjected to microarray expression analysis. The experiments were performed in triplicates.