Gene Expression
Nebulas
Gene Expression NebulasSummary: We sequenced the transcriptomes of seven samples of hypothalamic neurons dervied from pluripotent stem cells taken from healthy patients; five samples of hypothalamic neurons derived from pluripotent stem cells of constitutionally obese donors; five samples of sectioned hypothalami from post-mortem dissection of brains, and two samples for motor neurons derived from pluripotent stem cells of healthy donors in order to assess the similarity of our iPSC-derived cells against those of sectioned brains and to identify possible transcriptional disfunction which might underlie extreme, inherited obesity.
Overall Design: We sequenced mRNA collected from post-mortem brains and from samples generated by differentiating pluripotent stem cells into hypothalamic neurons and motor neurons.
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| Growth Protocol: | Induced Pluripotent Stem cell were differentiated into hypothalamic neurons for the iHT samples. IPSCs were differentiated into motor neurons for the iMN samples. Adult brains were dissected post-mortem to harvest natural hypothalamic neurons. |
| Treatment Protocol: | For differentiation into iHTNs, iPSCs were accutase-treated and plated as single cell in 6-well Matrigel-coated plates at a density of approx.; - |
| Extract Protocol: | RNA was extracted from all samples using the QIAGEN RNeasy mini kit. |
| Library Construction Protocol: | Libraries were constructed using the Illumina TruSeq Stranded mRNA library preparation kit. |
| Molecule Type: | poly(A)+ RNA |
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| Library Layout: | SINGLE |
| Library Strand: | Forward |
| Platform: | ILLUMINA |
| Instrument Model: | Illumina NextSeq 500 |
| Strand-Specific: | Specific |
| Data Resource | GEN Sample ID | GEN Dataset ID | Project ID | BioProject ID | Sample ID | Sample Name | BioSample ID | Sample Accession | Experiment Accession | Release Date | Submission Date | Update Date | Species | Race | Ethnicity | Age | Age Unit | Gender | Source Name | Tissue | Cell Type | Cell Subtype | Cell Line | Disease | Disease State | Development Stage | Mutation | Phenotype | Case Detail | Control Detail | Growth Protocol | Treatment Protocol | Extract Protocol | Library Construction Protocol | Molecule Type | Library Layout | Strand-Specific | Library Strand | Spike-In | Strategy | Platform | Instrument Model | Cell Number | Reads Number | Gbases | AvgSpotLen1 | AvgSpotLen2 | Uniq Mapping Rate | Multiple Mapping Rate | Coverage Rate |
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