Gut mucosa dissociation protocols influence cell type proportions and single-cell gene expression levels.

ABSTRACT: Single-cell RNA sequencing (scRNA-seq) has revolutionized the study of the cellular landscape of organs. Most single-cell protocols require fresh material, which limits sample size per experiment, and consequently, introduces batch effects. This is especially true for samples acquired through complex medical procedures, such as intestinal mucosal biopsies. Moreover, the tissue dissociation procedure required for obtaining single cells is a major source of noise; different dissociation procedures applied to different compartments of the tissue induce artificial gene expression differences between cell subsets. To overcome these challenges, we have developed a one-step dissociation protocol and demonstrated its use on cryopreserved gut mucosal biopsies. Using flow cytometry and scRNA-seq ana

SUBMITTER: Uniken Venema WTC 

PROVIDER: S-EPMC9197976 | biostudies-literature | 2022 Jun

REPOSITORIES: biostudies-literature