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Status |
Public on Dec 01, 2018 |
Title |
iPSC-NCCs (inflamed condition) 1 |
Sample type |
RNA |
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Source name |
coculture with a mix of PBMCs for 5 days
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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.
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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.
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Label |
biotin
|
Label protocol |
We followed protocol of manual that is GeneChip® WT PLUS Reagent Kit User Manual.
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Hybridization protocol |
We followed protocol of manual that is GeneChip® WT PLUS Reagent Kit User Manual.
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Scan protocol |
We followed manufacturer's protocol, and used GeneChip® Scanner 3000 7G.
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Data processing |
Data acquisition and normalization (SST-RMA) were used Affymetrix® Expression Console™ Software.
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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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