Functional heterogeneity of cancer-associated fibroblasts in pancreatic cancer

Single-cell RNA-sequencing analyses were performed on unfractionated live cell mixtures or sorted fibroblasts from pancreatic tumors of KPC;YFP mice, so as to investigate the functional heterogeneity of cancer-associated fibroblasts in early-stage and late-stage tumors. Overall design: Unfractionated live cell mixtures or sorted fibroblasts from pancreatic tumors of KPC;YFP mice were analyzed by 10x single-cell RNA-sequencing. Cancer-associated fibroblasts were extracted using the reverse sorting selection of CD45/CD31/YFP triple-negative cells. As an alternative route, aSMA-expressing cancer-associated fibroblasts were sorted by RFP expression from KPC;YFP;aSMA-RFP mice.

本研究针对KPC;YFP小鼠胰腺肿瘤来源的未经分选活细胞混合物或分选纯化的成纤维细胞开展单细胞RNA测序（single-cell RNA-sequencing）分析，旨在探究早、晚期肿瘤中癌症相关成纤维细胞（cancer-associated fibroblasts）的功能异质性。实验设计概况：采用10×单细胞RNA测序对KPC;YFP小鼠胰腺肿瘤来源的未经分选活细胞混合物或分选纯化的成纤维细胞进行检测分析。研究通过对CD45/CD31/YFP三阴性细胞实施反向分选筛选，分离得到癌症相关成纤维细胞；作为替代实验方案，可从KPC;YFP;aSMA-RFP小鼠中基于RFP表达特征分选得到表达α平滑肌肌动蛋白（aSMA）的癌症相关成纤维细胞。