Comparative analysis of 10X Chromium vs. BD Rhapsody whole transcriptome single-cell sequencing technologies in complex human tissues

10X Chromium 与 BD Rhapsody 全转录组单细胞测序技术在复杂人体组织中的比较分析

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作者:Stefan Salcher ,Isabel Heidegger ,Gerold Untergasser ,Georgios Fotakis ,Alexandra Scheiber ,Agnieszka Martowicz ,Asma Noureen ,Anne Krogsdam ,Christoph Schatz ,Georg Schäfer ,Zlatko Trajanoski ,Dominik Wolf ,Sieghart Sopper ,Andreas Pircher

Abstract

The development of single-cell omics tools has enabled scientists to study the tumor microenvironment (TME) in unprecedented detail. However, each of the different techniques may have its unique strengths and limitations. Here we directly compared two commercially available high-throughput single-cell RNA sequencing (scRNA-seq) technologies - droplet-based 10X Chromium vs. microwell-based BD Rhapsody - using paired samples from patients with localized prostate cancer (PCa) undergoing a radical prostatectomy. Although high technical consistency was observed in unraveling the whole transcriptome, the relative abundance of cell populations differed. Cells with low mRNA content such as T cells were underrepresented in the droplet-based system, at least partly due to lower RNA capture rates. In contrast, microwell-based scRNA-seq recovered less cells of epithelial origin. Moreover, we discovered platform-dependent variabilities in mRNA quantification and cell-type marker annotation. Overall, our study provides important information for selection of the appropriate scRNA-seq platform and for the interpretation of published results. Keywords: 10X Chromium; BD Rhapsody; Low-mRNA content cells; Neutrophils; Prostate cancer; Single-cell RNA sequencing.

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