Harvard Stem Cell Institute, Department of Stem Cell and Regenerative Biology, Harvard University, Cambridge, MA 02138, USA
Howard Hughes Medical Institute, Stanley Center for Psychiatric Research, Cambridge, MA 02142, USA.
Development. 2018 Nov 21;145(22):dev168617. doi: 10.1242/dev.168617.
Advances in stem cell science allow the production of different cell types either through the recapitulation of developmental processes, often termed 'directed differentiation', or the forced expression of lineage-specific transcription factors. Although cells produced by both approaches are increasingly used in translational applications, their quantitative similarity to their primary counterparts remains largely unresolved. To investigate the similarity between -derived and primary cell types, we harvested and purified mouse spinal motor neurons and compared them with motor neurons produced by transcription factor-mediated lineage conversion of fibroblasts or directed differentiation of pluripotent stem cells. To enable unbiased analysis of these motor neuron types and their cells of origin, we then subjected them to whole transcriptome and DNA methylome analysis by RNA sequencing (RNA-seq) and reduced representation bisulfite sequencing (RRBS). Despite major differences in methodology, lineage conversion and directed differentiation both produce cells that closely approximate the primary motor neuron state. However, we identify differences in Fas signaling, the Hox code and synaptic gene expression between lineage-converted and directed differentiation motor neurons that affect their utility in translational studies.
干细胞科学的进步使得人们可以通过重现发育过程（通常称为“定向分化”）或强制表达谱系特异性转录因子来产生不同的细胞类型。尽管这两种方法产生的细胞越来越多地应用于转化应用，但它们与原始细胞的定量相似性在很大程度上仍未得到解决。为了研究衍生细胞与原代细胞类型之间的相似性，我们收获并纯化了小鼠脊髓运动神经元，并将其与通过转录因子介导的成纤维细胞谱系转换或多能干细胞的定向分化产生的运动神经元进行了比较。为了能够对这些运动神经元类型及其起源细胞进行无偏分析，我们随后通过 RNA 测序（RNA-seq）和简化代表性双硫代测序（RRBS）对它们进行了全转录组和 DNA 甲基化组分析。尽管在方法上存在重大差异，但谱系转换和定向分化都产生了非常接近原始运动神经元状态的细胞。然而，我们发现谱系转换和定向分化产生的运动神经元在 Fas 信号、Hox 代码和突触基因表达方面存在差异，这影响了它们在转化研究中的应用。
