RNA-seq analysis of mouse pluripotent stem cells derived cells

To detect the role of OP9 stromal cells in our optimized 3D self-assembling peptide induction system followed by the OP9 coculture system. First, we used RNA-seq to analyze mouse pluripotent stem cells derived total cells at day5 between the 3D+OP9 and 3D+0.1% group in our hematopoietic differentiation system. In addition, to further evaluate hematopoietic transcriptome differences between Lin-Sca-1+c-kit+CD201+ cells and Lin-Sca-1+c-kit+CD201- cells, we used RNA-seq to analyze mouse pluripotent stem cells derived Lin-Sca-1+c-kit+CD201+ and Lin-Sca-1+c-kit+CD201- cells at day5 in our 3D+OP9 hematopoietic differentiation system.Meanwhile,we used the mouse embryonic stem cell(mESC) as negative control and bone marrow derived Lin-Sca-1+c-kit+CD201+ and fetal liver derived Lin-Sca-1+c-kit+CD201+ as positive control. Overall design: two groups together: 3 sample of 3D+OP9 and 3 sample of 3D+0.1%gel; five groups together : 3 sample of D5-LSKCD201+ 3 sample of D5-LSKCD201- 3 sample of mESC 3 sample of BM-LSKCD201+ 3 sample of FL-LSKCD201+

本研究旨在探究OP9基质细胞（OP9 stromal cells）在本实验室优化的3D自组装肽诱导体系联合OP9共培养体系中所发挥的作用。首先，我们采用RNA测序（RNA-seq）技术，对造血分化体系中第5天的3D+OP9组与3D+0.1%凝胶组的小鼠多能干细胞来源总细胞进行转录组比较分析。此外，为进一步明确Lin-Sca-1+c-kit+CD201+细胞与Lin-Sca-1+c-kit+CD201-细胞之间的造血转录组差异，我们通过RNA-seq技术，分析了3D+OP9造血分化体系中第5天的小鼠多能干细胞来源的Lin-Sca-1+c-kit+CD201+与Lin-Sca-1+c-kit+CD201-细胞。与此同时，我们以小鼠胚胎干细胞（mouse embryonic stem cell, mESC）作为阴性对照，以骨髓来源的Lin-Sca-1+c-kit+CD201+细胞与胎肝来源的Lin-Sca-1+c-kit+CD201+细胞作为阳性对照。整体实验设计分为两部分：第一部分包含2组样本，分别为3份3D+OP9组样本与3份3D+0.1%凝胶组样本；第二部分包含5组样本，分别为3份第5天LSKCD201+组样本、3份第5天LSKCD201-组样本、3份小鼠胚胎干细胞组样本、3份骨髓来源LSKCD201+组样本以及3份胎肝来源LSKCD201+组样本。