概要: 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.
项目整体设计: We sequenced mRNA collected from post-mortem brains and from samples generated by differentiating pluripotent stem cells into hypothalamic neurons and motor neurons.
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.
处理方案:
For differentiation into iHTNs, iPSCs were accutase-treated and plated as single cell in 6-well Matrigel-coated plates at a density of approx.; -
提取方案:
RNA was extracted from all samples using the QIAGEN RNeasy mini kit.
建库方案:
Libraries were constructed using the Illumina TruSeq Stranded mRNA library preparation kit.
