Comment[ArrayExpressAccession] E-MEXP-3986
Investigation Title MiRNA expression in airway epithelial wound repair_Szczepankiewicz
Comment[Submitted Name] MiRNA expression in airway epithelial wound repair_Szczepankiewicz
Comment[AEExperimentDisplayName] MicroRNA profiling by array of human bronchial epithelial cells after wounding
Comment[MIAMExpressLogin] alszczep
Comment[MIAMExpressSubmissionID] 8900
Experiment Description Analysis of miRNA expression at different time points of epithelial wound repair of 16HBE14o- cells after mechanical damage of cell monolayer.
Experimental Design in_vitro_design co-expression_design injury design time series design
Comment[AEExperimentType] microRNA profiling by array
Experimental Factor Name time
Experimental Factor Type time
Person Last Name Szczepankiewicz
Person First Name Aleksandra
Person Email alszczep@gmail.com
Person Phone +48 61-8491311
Person Affiliation Poznan University of Medical Sciences
Person Address Laboratory of Molecular and Cell Biology, 27/33 Szpitalna St., Poznan, Wielkopolska, 60-572, Poland
Person Roles submitter
Person Roles Term Source REF EFO
Quality Control Type technical replicate
Comment[QualityControlDescription] For each time point 3 technical replicates were used.
Public Release Date 2013-10-02
Comment[ArrayExpressSubmissionDate] 2013-10-02 22:15:30
Publication Title Altered microRNA expression profile during epithelial wound repair in bronchial epithelial cells
Publication Status in press
Protocol Name P-MTAB-34995 P-MTAB-34996 P-MTAB-34997 P-MTAB-34997 P-MTAB-34998 P-MTAB-34999
Protocol Description For the wounding assay cells were seeded on 6-well plates at the initial density of 3x105 cells and cultured until confluent. Forty eight hours after reaching full confluence cells were damaged by scraping off the monolayer with a hatch-cross wounding pattern using a P200 Gilson pipette tip. After that, the medium and cell debris were removed and 2 ml of fresh serum-containing medium was added to the remaining cells. For all experiments, at least two points of reference per well of a 6-well plate were used for post-injury analyses. For miRNA expression profile 7 time points were selected. Labeling was done by Applied Biosystems.
(Parameters: Amplification = PCR, Mass unit = Micro gram) The 16HBE14o- bronchial epithelial cell line was cultured under standard conditions described by Adam et al. (2007).
(Parameters: time unit = seconds, min temperature = 37, temperature unit = C, media = MEM (Gibco)) Hybridization was done by Applied Biosystems.
(Parameters: Chamber type = OTHER: TaqMan Human miRNA cards A and B, Quantity of label target used = 0, Mass unit = Micro gram, Tiny time unit = seconds, Volume unit = Nano litre, temperature = 0) RNA isolation was performed with use of Exiqon RNA isolation kit. Isolation was performed according to the manufacturerM-^Rs protocol from 6-well plates (9.5 cm2 of growth area) and the amount of starting material was 1x106 cells per well. Samples were frozen at -70M-0C for further use in microarray experiment.
(Parameters: Extracted product = total_RNA, Amplification = PCR) This step was done according to manufacturer's (Applied Biosystems) instructions.
(Parameters: Scanning hardware = OTHER: AbiPrism 7900 HT, Scanning software = Scanning software)
Protocol Type treatment protocol nucleic acid labeling protocol growth protocol nucleic acid hybridization to array protocol nucleic acid extraction protocol array scanning protocol
Protocol Term Source REF EFO EFO EFO EFO EFO EFO
Term Source Name EFO
Term Source File http://www.ebi.ac.uk/efo/
SDRF File E-MEXP-3986.sdrf.txt