A single-cell RNA-sequencing time course of liver colonisation by colorectal cancer metastases. A single-cell RNA-sequencing time course of liver colonisation by colorectal cancer metastases

Murine colorectal cancer cells were injected into the portal vein of NOD.Cg-PrkdcSCID Il2rgtm1Wjl/SzJ mice. Liver metastatic cells were harvested at specific time points for single-cell RNA-sequencing (SORT-seq). Overall design: Lgr5- or Lgr5+ cells were collected from RFP+ murine colorectal cancer organoids (genetic background VillinCre-ERT2; ApcFL/FL; KrasLSL-G12D/+; Tp53KO/KO; R26R-Confetti; Lgr5DTR-eGFP) by FACS and injected into the portal vein of recipient mice. After 1, 7, 13, and 27 days, livers were perfused with a collagenase cocktail and RFP+ metastatic cells were isolated by FACS and analysed by single-cell RNA-seq.

将小鼠结直肠癌细胞注射至NOD.Cg-PrkdcSCID Il2rgtm1Wjl/SzJ小鼠的门静脉内。于特定时间点收集肝转移细胞，用于单细胞RNA测序（single-cell RNA-sequencing，SORT-seq）。
实验设计：从遗传背景为VillinCre-ERT2; ApcFL/FL; KrasLSL-G12D/+; Tp53KO/KO; R26R-Confetti; Lgr5DTR-eGFP的RFP+小鼠结直肠癌类器官中，通过荧光激活细胞分选（fluorescence-activated cell sorting，FACS）分选出Lgr5阴性（Lgr5-）或Lgr5阳性（Lgr5+）细胞，并将其注射至受体小鼠的门静脉内。分别于细胞注射后1天、7天、13天和27天，采用胶原酶混合液对小鼠肝脏进行灌注，随后通过荧光激活细胞分选（FACS）分离RFP+转移细胞，再进行单细胞RNA测序分析。