scRNAseq of Cyp11a1-expressing cells from mouse melanoma model derived tumors

Tumours induce de novo steroidogenesis in immune cells. We wanted to define the gene expression identity of intratumoural steroidogenic immune cells and compare them with the transcriptome with non-steroidogenic cells. Cyp11a1 expression is a faithful biomarker of de novo steroidogenesis (i.e. Cyp11a1+ cells are steroidogenic and Cyp11a1- cells are non-steroidogenic). We inoculated B16-F10 tumor in Cyp11a1-mCherry reporter mice, enriched and purified intratumoral Cyp11a1-mCherry+ and Cyp11a1-mCherry- cells by cell sorting into 96-well plates [with a ratio of 79:15 (mCherry+ : mCherry-) cells per plate] and performed scRNA-seq using SMART-Seq2 platform.

肿瘤可诱导免疫细胞发生从头类固醇生成（de novo steroidogenesis）。本研究旨在明确肿瘤内类固醇生成性免疫细胞的基因表达特征，并将其与非类固醇生成性免疫细胞的转录组进行对比。细胞色素P450 11A1（Cyp11a1）的表达是从头类固醇生成的可靠生物标志物，即Cyp11a1阳性（Cyp11a1+）细胞为类固醇生成性细胞，Cyp11a1阴性（Cyp11a1-）细胞为非类固醇生成性细胞。我们向Cyp11a1-mCherry报告基因小鼠体内接种B16-F10肿瘤细胞，通过细胞分选将肿瘤内的Cyp11a1-mCherry阳性与阴性细胞富集纯化后接种至96孔板中，每板的细胞比例为79:15（mCherry阳性: mCherry阴性），并采用SMART-Seq2平台进行单细胞RNA测序（single-cell RNA sequencing, scRNA-seq）。