|
| Status |
Public on Nov 28, 2018 |
| Title |
Cynomolgus monkey PBMCs treated with LPS (2 μg.ml-1) replicate 2 |
| Sample type |
RNA |
| |
|
| Channel 1 |
| Source name |
The PBMCs (1×106 cells/ml) obtained from the cynomolgus monkeys were treated with LPS ( 2 μg.ml-1 ) and incubated at 37 °C for 3 days, replicate 2
|
| Organism |
Macaca fascicularis |
| Characteristics |
tissue: peripheral blood
cell type: mononuclear
process: test
stimulation: LPS
|
| Treatment protocol |
The PBMCs (1×10^6 cells/ml) obtained from the cynomolgus monkeys or the human donors were treated with PHA (2 μg.ml-1 or 10 μg.ml-1) or LPS (2 μg.ml-1 or 10 μg.ml-1) and incubated at 37 °C for 3 days. Cells cultured under similar conditions without any stimulation served as the negative control.
|
| Growth protocol |
Whole heparinized cynomolgus monkey blood was obtained from 3 apparently healthy 2.5-year-old cynomolgus monkeys. The monkey PBMCs were isolated by Ficoll-Hypaque density centrifugation (Histopaque LTS1079, Sigma, USA). All procedures were performed in accordance with the Guide for the Care and Use of Laboratory Animals of NCSED, China. A relative centrifugal force of 2000 rpm was used for 30 minutes to separate the PBMCs. The PBMCs were washed twice in Hank’s and re-suspended at a final concentration of 1×10^6 cells/ml in culture medium (RPMI1640, GIBCO, USA) supplemented with 10% fetal calf serum. All experiments using the cultured cells were set up in triplicate.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
The total RNA was extracted from the PBMCs using Trizol reagent (Invitrogen Company) according to the manufacturer’s instructions. The purity of the RNA in each sample was assessed spectrophotometerically at 260 and 280 nm (NanoDrop Technologies), and the RNA integrity was assessed using formaldehyde agarose gel electrophoresis.
|
| Label |
Cy5
|
| Label protocol |
not provided
|
| |
|
| Channel 2 |
| Source name |
The PBMCs cultured under similar conditions without any stimulation served as the negative control, replicate 2
|
| Organism |
Macaca fascicularis |
| Characteristics |
tissue: peripheral blood
cell type: mononuclear
process: control
|
| Treatment protocol |
The PBMCs (1×10^6 cells/ml) obtained from the cynomolgus monkeys or the human donors were treated with PHA (2 μg.ml-1 or 10 μg.ml-1) or LPS (2 μg.ml-1 or 10 μg.ml-1) and incubated at 37 °C for 3 days. Cells cultured under similar conditions without any stimulation served as the negative control.
|
| Growth protocol |
Whole heparinized cynomolgus monkey blood was obtained from 3 apparently healthy 2.5-year-old cynomolgus monkeys. The monkey PBMCs were isolated by Ficoll-Hypaque density centrifugation (Histopaque LTS1079, Sigma, USA). All procedures were performed in accordance with the Guide for the Care and Use of Laboratory Animals of NCSED, China. A relative centrifugal force of 2000 rpm was used for 30 minutes to separate the PBMCs. The PBMCs were washed twice in Hank’s and re-suspended at a final concentration of 1×10^6 cells/ml in culture medium (RPMI1640, GIBCO, USA) supplemented with 10% fetal calf serum. All experiments using the cultured cells were set up in triplicate.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
The total RNA was extracted from the PBMCs using Trizol reagent (Invitrogen Company) according to the manufacturer’s instructions. The purity of the RNA in each sample was assessed spectrophotometerically at 260 and 280 nm (NanoDrop Technologies), and the RNA integrity was assessed using formaldehyde agarose gel electrophoresis.
|
| Label |
Cy3
|
| Label protocol |
not provided
|
| |
|
| |
| Hybridization protocol |
The purified total RNA was amplified and labeled, and the microarray analysis was then performed overnight using Agilent Human and Monkey Whole Genome GeneChips (Agilent, Palo Alto, CA).
|
| Scan protocol |
not provided
the array data were analyzed using Agilent Feature Extraction software.
|
| Description |
Cy5/Cy3
|
| Data processing |
After normalization of the raw data, probes with intensities <400 were removed from the subsequent analysis using Significance Analysis of Microarray (SAM) software. The differentially expressed genes (DEGs) were identified as those having an adjusted q-value of <0.05 and an average fold-change difference of >1.5.
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| |
|
| Submission date |
Nov 30, 2016 |
| Last update date |
Nov 28, 2018 |
| Contact name |
wang liang |
| E-mail(s) |
wangliangkexue@163.com
|
| Organization name |
BioChainBJ
|
| Street address |
7A North Yongchang Road BDA
|
| City |
Beijing |
| ZIP/Postal code |
100176 |
| Country |
China |
| |
|
| Platform ID |
GPL22725 |
| Series (2) |
| GSE90722 |
The morphological and functional differences and similarities of immune cells between non-human primates and humans [monkey] |
| GSE90723 |
The morphological and functional differences and similarities of immune cells between non-human primates and humans |
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