Single-cell RNA-Sequencing maps the hematopoietic landscape in naked mole-rats [BM]

Naked mole-rats are a mammalian model organism of exceptional longevity. We mapped the hematopoietic hierarchy and used functional characterizations to define a purified stem and progenitor cell (HSPC) compartment similar to mouse LIN-/Sca-1+/Kit+ HSPCs or human LIN-/CD34+/CD38lo HSPCs. Naked mole-rat or Mouse bone marrow was extracted, RBC-lysed and submitted to scRNA-Seq as unfractionated BM, in parallel FACS-antibodies were conjugated to oligo-ADT tags to quantify surface marker binding as described for CITE-Seq. The mRNA libraries were aligned to either the FRAMA bone marrow transcriptome assembly from our publication or a mouse genome reference.裸鼹鼠是一类具备超长寿命的哺乳动物模式生物。本研究绘制了其造血层级图谱，并通过功能表征实验定义了一类纯化的造血干祖细胞（hematopoietic stem and progenitor cell, HSPC）群体，该群体与小鼠LIN-/Sca-1+/Kit+型造血干祖细胞或人类LIN-/CD34+/CD38lo型造血干祖细胞特征相似。分别提取裸鼹鼠与小鼠的骨髓（bone marrow, BM）组织，经红细胞裂解处理后，将未分选的骨髓样本提交至单细胞RNA测序（single-cell RNA sequencing, scRNA-Seq）流程；同时参照转录组与表位细胞索引测序（Cellular Indexing of Transcriptomes and Epitopes by Sequencing, CITE-Seq）的实验方案，将荧光激活细胞分选（fluorescence-activated cell sorting, FACS）抗体与寡核苷酸ADT标签偶联，以量化表面标记物的结合水平。所构建的mRNA文库分别比对至本团队已发表研究中的FRAMA骨髓转录组组装集，或小鼠基因组参考序列。