|
| Status |
Public on Jan 07, 2017 |
| Title |
PIE cells with challenge |
| Sample type |
RNA |
| |
|
| Source name |
PIE cells, no challenge, replicate 1
|
| Organism |
Sus scrofa domesticus |
| Characteristics |
cell line: Porcine intestinal epitheliocytes-derived cell line
age: 3 days
|
| Growth protocol |
PIE cells, which are non-transformed intestinal cultured cells originally derived from intestinal epithelia isolated from an unsuckled neonatal swine, were maintained in Dulbecco’s modified Eagle’s medium (DMEM) (Invitrogen Corporation, Carlsbad, CA) supplemented with 10% fetal calf serum (FCS), 100 mg/ml penicillin, and 100 U/ml streptomycin at 37°C in an atmosphere of 5% CO2. PIE cells were seeded at 3 x 104 cells/12-well plate on type I collagen-coated plates (Sumitomo Bakelite Co., Tokyo, Japan) and cultured for 3 days. Control medium or lactobacilli (5 x 108 cells/ml) were added and 48 h later, cells were stimulated with Poly(I:C) (60 ng/ml). 12 h after Poly(I:C) stimulation total RNA was extracted for microarray studies.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was isolated from PIE cells samples using PureLink RNA Mini Kit (life technologies inc. gaithersburg md) with DNase. RNA integrity of all samples were evaluated by Agilent 2100 Bioanalyzer (Agilent Technologies, Santa Clara, CA, USA), using the RNA 6000 Nano Kit.
|
| Label |
Cy3
|
| Label protocol |
Labelling was done as recommended by Agilent Technologies using 200 ng of RNA
|
| |
|
| Hybridization protocol |
Hybridisation with Porcine (V2) Gene Expression Microarray (Agilent Technologies) was performed at Hokkaido System Science Co.
|
| Scan protocol |
Scanned on an Agilent G2505C scanner.
Images were quantified using Agilent Feature Extraction Software (version 10.7.3.1).
|
| Description |
Gene expression after 12h Poly(I:C) challenge
|
| Data processing |
Data normalization and expression analysis were performed with GeneSpring software version 6.1 (Agilent Technologies).
Expression data were considered significant when they differed at least 2 fold between treated and untreated PIE cells, and results are expressed as log2 scale (log2 ratio). Genes whose expressions were increased more than 2-fold (log2 > 1) after treatment with Poly(I:C) were selected
|
| |
|
| Submission date |
Jan 06, 2017 |
| Last update date |
Jan 07, 2017 |
| Contact name |
Julio Villena |
| E-mail(s) |
jcvillena@cerela.org.ar
|
| Phone |
(+54) (381) 4310465
|
| Organization name |
CERELA-CONICET
|
| Lab |
Laboratory of Immunobiotechnology
|
| Street address |
Chacabuco 145
|
| City |
San Miguel de Tucumán |
| State/province |
Tucumán |
| ZIP/Postal code |
4000 |
| Country |
Argentina |
| |
|
| Platform ID |
GPL15007 |
| Series (1) |
| GSE93225 |
PIE cell line: Control vs. Poly(I:C) treated or Immunobiotic treated |
|
