外周血单个核细胞
生物
分子生物学
小瓶
免疫系统
基因表达
核糖核酸
单元格排序
背景(考古学)
细胞
基因
免疫学
遗传学
流式细胞术
化学
体外
古生物学
物理化学
作者
Gloria Kraus,Marc Weigelt,Susanne Reinhardt,A. Petzold,Andreas Dahl,Ezio Bonifacio
标识
DOI:10.1016/j.jim.2022.113227
摘要
10x Genomics is a highly accessible single cell RNA sequencing platform that allows for simultaneous gene expression analysis and identification of receptor chain combinations in cells of the adaptive immune system. Here, we asked whether the gene and receptor expression measurements in peripheral blood mononuclear cells (PBMC) are influenced by technical, cell freezing, FACS-processing, and day to day biological variation. No differentially expressed gene was observed between 1. triplicates aliquots taken from the same vial of frozen PBMC; 2. triplicate vials of frozen PBMC; and 3. triplicate aliquots taken from the same vial of frozen PBMC and processed separately for FACS staining and sorting of different PBMC populations. A small number of differentially expressed genes were observed between PBMC sampled, isolated and frozen from the same donor on different days, and these differences were more pronounced in the memory B cells than other cell populations. T cell receptors were recovered in all replicates when at least 5 cells per clonotype were identified. These findings show high reproducibility of 10x Genomics single cell RNA sequencing data in the immune cell context.
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