10X-single cell seq of B cells during colitis (d0 and d14). 10X-single cell seq of B cells during colitis (d0 and d14)

B cells expand during the recovery after DSS-induced colonic inflammation. To investigate if a specific subtype is expanding and to analyse the transcriptional profile of these cells a transcriptional analysis on single cell level was carried out. In detail, B cells were isolated from the colon of mice on day 0 (no treatment) or day 14 (recovery) after 7 days of DSS treatment by flow cytometric sorting and analyzed by 10X sequencing. Overall design: Female C57BL/6J mice were treated with 2%DSS via the drinking water followed by 7 days of recovery. Lymphocytes were isolated from the colonic lamina propria on day 0 (ctrl) and day 14 (recovery) and B cells were sorted using flow cytometry. Afterwards cells were processed according to 10x Genomics protocol for Chromium Single cell 3’ kit (version 2) and submitted for sequencing

B细胞在葡聚糖硫酸钠（Dextran Sulfate Sodium, DSS）诱导的结肠炎症恢复阶段发生扩增。为明确是否存在特定B细胞亚型发生扩增，并解析此类细胞的转录组特征，本研究开展了单细胞水平的转录组分析。具体而言，研究人员分别于第0天（未处理对照组）及第14天（恢复阶段），从经7天DSS处理后的小鼠结肠中分离B细胞，经流式细胞术分选后采用10X测序技术进行分析。
实验总体设计：选取雌性C57BL/6J小鼠，通过饮用水给予2%DSS处理，随后进行7天恢复。分别于第0天（对照组）及第14天（恢复阶段）从结肠固有层分离淋巴细胞，经流式细胞术分选出B细胞。后续细胞样本按照10x Genomics的Chromium单细胞3’端试剂盒（版本2）标准操作流程进行制备，并送交测序。