|
| Status |
Public on Dec 01, 2018 |
| Title |
iPSC-NCCs (normal condition) 2 |
| Sample type |
RNA |
| |
|
| Source name |
monoculture for 5 days
|
| Organism |
Homo sapiens |
| Characteristics |
cell line: 253G1
cell type: neural crest cell
|
| Growth protocol |
We induced NCCs from human iPSCs according to the previously described method with some modifications for 3 weeks to complete. After the differentiation, iPSC-NCCs were harvested in a 12-well culture plate. The medium was RPMI 1640 (Nacalai Tesque) containing 10% FBS supplemented with human recombinant interleulin-2 (BD). iPSC-NCCs (normal condition, 1.0 x 10^6 cells/well) were incubated in the absence of the transwell membrane. On the other hand, iPSC-NCCs (inflamed condition, 1.0 x 10^6 cells/well) were incubated in the presence of the transwell membrane. A mix of the human peripheral blood mononuclear cells (PBMCs, 3.0 x 10^6 cells/well) from 5 healthy donors were added to the upper wells of a transwell plate. PBMCs were obtained Lymphoprep (Stemcell technologies) density gradient centrifugation in accordance with the manufacturer’s instructions.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
The total RNA of iPSC-NCCs (normal condition, n=2) and iPSC-NCCs (inflamed condition, n=2) were extracted by High Pure RNA Isolation Kit (Roche) after the cells were incubated for 5 days.
|
| Label |
biotin
|
| Label protocol |
We followed protocol of manual that is GeneChip® WT PLUS Reagent Kit User Manual.
|
| |
|
| Hybridization protocol |
We followed protocol of manual that is GeneChip® WT PLUS Reagent Kit User Manual.
|
| Scan protocol |
We followed manufacturer's protocol, and used GeneChip® Scanner 3000 7G.
|
| Data processing |
Data acquisition and normalization (SST-RMA) were used Affymetrix® Expression Console™ Software.
|
| |
|
| Submission date |
Jun 21, 2018 |
| Last update date |
Dec 02, 2018 |
| Contact name |
Shota Fujii |
| Organization name |
RIKEN Center for Developmental Biology
|
| Department |
The Laboratory for Retinal Regeneration
|
| Street address |
2-2-3 Minatojima-minamimachi, Chuo-ku
|
| City |
Kobe |
| ZIP/Postal code |
650-0047 |
| Country |
Japan |
| |
|
| Platform ID |
GPL23159 |
| Series (1) |
| GSE116110 |
Immunological properties of neural crest cells derived from human induced pluripotent stem cells (normal vs inflammed condition) |
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