Single-Cell RNA-seq study of E13.5 and P7 brain and Spinal Cord Pdgfra-GFP positive samples during development in Mice.

In order to elucidate the developmental origin of oligodendrocyte precursor cells (OPCs) and get a better understanding of the several waves of OPC generation, we look at several timepoints and perform single-cell RNA-seq on Pdgfra positive populations in Mice. Overall design: Mice line used in this study included Pdgfra-cre-ERT/RCE and the Pdgfra-H2BGFP knock-in mouse. Embryos at embryonic day 13.5 and pups from post-natal day 7, from both genders of the Pdgfra-GFP mice line were used to extract OPCs, as well as E12.5 and P3 tamoxifen injected mice harvested at P7. The single cell suspension from embryonic and post-natal tissue was FACS sorted for GFP positive cells using a BD FACSAria III Cell Sorter B5/R3/V3 system.为阐明少突胶质前体细胞（oligodendrocyte precursor cells, OPCs）的发育起源，并深入理解少突胶质前体细胞生成的多波次过程，本研究选取多个时间节点，对小鼠体内Pdgfra阳性细胞群开展单细胞RNA测序（single-cell RNA-seq）。 总体实验设计：本研究使用的小鼠品系包括Pdgfra-cre-ERT/RCE品系与Pdgfra-H2BGFP敲入小鼠。实验取材对象涵盖：Pdgfra-GFP小鼠品系的13.5天胚胎（embryonic day 13.5, E13.5）与产后7天（post-natal day 7, P7）的雌雄个体，以及于胚胎12.5天（E12.5）或产后3天（post-natal day 3, P3）注射他莫昔芬、并于产后7天（P7）取材的小鼠。从胚胎及产后组织制备单细胞悬液后，采用BD FACSAria III细胞分选仪（BD FACSAria III Cell Sorter B5/R3/V3）分选出GFP阳性细胞。