Single-cell RNA sequencing analysis of non-haematopoietic DNGR-1 lineage traced cells from murine spinal cord

Stem cells are defined by two cardinal properties: limitless self-renewal and multipotency. We have serendipitously found that non-haematopoietic DNGR-1 lineage traced cells residing in the ependymal cell layer of the central nervous system display the two cardinal properties of stem cells, both in vitro and in vivo. However, whether these properties were a feature of all DNGR-1-traced cells or were confined to a particular subset of these is unclear. To address the potential heterogeneity of DNGR-1-traced ependymal cells and characterize their putative stem cell compartment we conducted single-cell RNA sequencing of DNGR-1-traced cells isolated from uninjured spinal cords. DNGR-1 lineage traced cells pooled from a spinal cords dissected from DNGR-1 lineage tracer mice were FACS sorted and subjected to single-cell RNA sequencing干细胞的定义以两项核心特征为依据：无限自我更新能力与多向分化潜能。我们意外发现，栖息于中枢神经系统室管膜层的非造血系DNGR-1谱系示踪细胞，在体外与体内环境中均展现出干细胞的这两项标志性特征。然而目前仍不清楚，这些特征是所有DNGR-1谱系示踪细胞共有的属性，还是仅局限于该细胞群的某一特定亚群。为探究DNGR-1谱系示踪室管膜细胞的潜在异质性，并对其推定的干细胞亚群进行表征，我们对从未损伤脊髓中分离得到的DNGR-1谱系示踪细胞开展了单细胞RNA测序（single-cell RNA sequencing）。从DNGR-1谱系示踪小鼠体内解剖获取的脊髓中收集的DNGR-1谱系示踪细胞，经荧光激活细胞分选（FACS）富集后，进行了单细胞RNA测序。