|
| Status |
Public on Apr 01, 2024 |
| Title |
Monocytes THP-1: Control vs Colpvp |
| Sample type |
RNA |
| |
|
| Channel 1 |
| Source name |
Total RNA extracted from untreated monocytes
|
| Organism |
Homo sapiens |
| Characteristics |
cell line: THP-1
cell type: Macrophage
|
| Treatment protocol |
Controls were untreated cell cultures, Colpvp were incubated for 3 h with FBS-free medium and then with 3% collagen-PVP prepared in the corresponding medium without FBS and incubated for 24 h at 37 °C and 5% CO2
|
| Growth protocol |
The human monocyte cell line (THP-1) was cultured with RPMI-1640 medium, supplemented with 10% FBS, 1mM sodium pyruvate, 0.1mM non-essential aminoacids, 0.1mM of glutamine, 100 U / ml penicillin and 100 μg/ml streptomycin.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA extracted using Trizol following manufacturer's instructions
|
| Label |
Cy3
|
| Label protocol |
10 µg of total RNA for cDNA synthesis, incorporating dUTP-Alexa555 or dUTP-Alexa647 using the First-Strand cDNA labeling kit (Invitrogen). Incorporation of fluorophore was analyzed by using the absorbance at 555 or 650 nm.
|
| |
|
| Channel 2 |
| Source name |
Total RNA extracted from Colpvp treated monocytes
|
| Organism |
Homo sapiens |
| Characteristics |
cell line: THP-1
cell type: Macrophage
|
| Treatment protocol |
Controls were untreated cell cultures, Colpvp were incubated for 3 h with FBS-free medium and then with 3% collagen-PVP prepared in the corresponding medium without FBS and incubated for 24 h at 37 °C and 5% CO2
|
| Growth protocol |
The human monocyte cell line (THP-1) was cultured with RPMI-1640 medium, supplemented with 10% FBS, 1mM sodium pyruvate, 0.1mM non-essential aminoacids, 0.1mM of glutamine, 100 U / ml penicillin and 100 μg/ml streptomycin.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA extracted using Trizol following manufacturer's instructions
|
| Label |
Cy5
|
| Label protocol |
10 µg of total RNA for cDNA synthesis, incorporating dUTP-Alexa555 or dUTP-Alexa647 using the First-Strand cDNA labeling kit (Invitrogen). Incorporation of fluorophore was analyzed by using the absorbance at 555 or 650 nm.
|
| |
|
| |
| Hybridization protocol |
Hybridization was performed using equal amounts of labeled cDNA with the hybridization solution UniHyb (TeleChem International INC). The arrays were incubated ( 42°C, 14 hrs), and washed with 1X SCC/0.05 % SDS at RT.
|
| Scan protocol |
GenePix 4100A and GenePix software were used for the acquisition and quantification of array images using 10µm resolution
|
| Description |
Biological sample 1 of 2, untreated, macrophages
Biological sample 2 of 2, colpvp, macrophages
|
| Data processing |
ArrayPro Analyzer (Media Cibernetics) was used to calculate the Alexa555 and Alexa647 density mean value and background mean value for each spot. genArise (http://www.ifc.unam.mx/genarise/) was used for the microarray data performing background correction, lowess normalization, intensity filter, replicates analysis and selecting differentially expressed genes.
|
| |
|
| Submission date |
Mar 28, 2024 |
| Last update date |
Apr 01, 2024 |
| Contact name |
Krötzsch Edgar |
| E-mail(s) |
kroted@yahoo.com.mx
|
| Organization name |
Instituto Nacional de Rehabilitación LGII
|
| Lab |
Tejido Conjuntivo
|
| Street address |
Calz México-Xochimilco 289, Coapa, Col. Arenal de Guadalupe, Tlalpan
|
| City |
Mexico City |
| ZIP/Postal code |
14389 |
| Country |
Mexico |
| |
|
| Platform ID |
GPL34346 |
| Series (1) |
| GSE262737 |
Macrophages THP-1: Control vs Colpvp |
|
