Genome-wide analysis of transcription in in vitro- and in vivo-derived motor neurons. Mus musculus

RNA sequencing analysis of Hb9::GFP mouse embryonic fibroblasts, Hb9::GFP+ primary mouse embryonic motor neurons at day E13.5, Hb9::GFP+ mouse embryonic stem cell-derived motor neurons, Hb9::GFP+ mouse induced pluripotent stem cell derived motor neurons, and Hb9::GFP+ mouse induced motor neurons generated using transcription factor overexpression. The goal of this project is to evaluate the ability of directed differentiation and lineage conversion techniques to generate a bona fide neuronal subtype. Overall design: The goal of this study is to determine the level of transcriptional similarity between spinal motor neurons generated from pluripotent stem cells by directed differentation, spinal motor neurons generated from fibroblasts by transcription factor-mediated lineage conversion, and primary spinal motor neuron from embryonic spinal cord. In addition, we tested the effect of culturing ESC-MNs in glia-conditioned media. All neurons were generated using the same Hb9::GFP, C57Bl/6 background, and FACS sorted for Hb9::GFP.

本数据集为针对以下样本的RNA测序分析结果：Hb9::GFP标记的小鼠胚胎成纤维细胞、胚胎第13.5天（E13.5）的Hb9::GFP阳性原代小鼠胚胎运动神经元、Hb9::GFP阳性小鼠胚胎干细胞来源运动神经元、Hb9::GFP阳性小鼠诱导多能干细胞来源运动神经元，以及通过转录因子过表达制备的Hb9::GFP阳性小鼠诱导运动神经元。本项目的研究目标为评估定向分化与谱系重编程技术生成纯正神经元亚型的能力。本实验的整体设计如下：旨在明确三类脊髓运动神经元的转录相似性水平——通过定向分化从多能干细胞获得的脊髓运动神经元、通过转录因子介导的谱系重编程从成纤维细胞获得的脊髓运动神经元，以及胚胎脊髓来源的原代脊髓运动神经元。此外，本研究还检测了在胶质细胞条件培养基中培养胚胎干细胞来源运动神经元（ESC-MNs）的效果。所有受试神经元均采用统一的Hb9::GFP、C57Bl/6背景品系，并通过荧光激活细胞分选（Fluorescence Activated Cell Sorting, FACS）筛选Hb9::GFP阳性细胞。