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