Single cell RNAseq reveal covid19 patient immune response

Eighteen patients admitted to CSMC and diagnosed with COVID19 by RT-PCR were stratified into COVID19 mild/moderate, severe, and recovery groups (n=5-6/group). Venous blood was collected into EDTA coated tubes and centrifuged to separate plasma and buffy coat. Plasma was collected and frozen at -80C, and the buffy coat was collected into cryo-preservation media and frozen at -80C. Recovered cells are sorted for live-dead staining, then fixed with methanol. Fixed single cells were further captured using 10x chromium Next GEM 3 prime v3.1 kit. Two patients samples from the same group were mixed, captured and sequenced together. Overall design: single-cell RNA-seq of blood buffy coat from COVID-19 patients.

18名收入CSMC医院、经逆转录聚合酶链反应（RT-PCR）确诊为新型冠状病毒肺炎（COVID-19）的患者，按病情分层为轻症/普通型新冠组、重症新冠组及康复组（每组n=5~6例）。采集患者静脉血至乙二胺四乙酸（EDTA）抗凝管中，经离心分离得到血浆与棕黄层（buffy coat）；分离所得血浆被收集并置于-80℃冻存，棕黄层则转移至低温保存液中，同样于-80℃冻存。复苏后的细胞先通过细胞死活染色进行分选，随后用甲醇固定。固定后的单个细胞采用10x Chromium Next GEM 3 prime v3.1试剂盒（10x chromium Next GEM 3 prime v3.1 kit）进行捕获。同一组的两名患者的样本将被混合后，一同进行捕获与测序。本实验整体设计为：对新冠患者血液棕黄层开展单细胞RNA测序（single-cell RNA-seq）。