Enzymatic Dissociation Induces Transcriptional and Proteotype Bias in Brain Cell Populations.

ABSTRACT: Different cell isolation techniques exist for transcriptomic and proteotype profiling of brain cells. Here, we provide a systematic investigation of the influence of different cell isolation protocols on transcriptional and proteotype profiles in mouse brain tissue by taking into account single-cell transcriptomics of brain cells, proteotypes of microglia and astrocytes, and flow cytometric analysis of microglia. We show that standard enzymatic digestion of brain tissue at 37 °C induces profound and consistent alterations in the transcriptome and proteotype of neuronal and glial cells, as compared to an optimized mechanical dissociation protocol at 4 °C. These findings emphasize the risk of introducing technical biases and biological artifacts when implementing enzymatic digestion-based is

SUBMITTER: Mattei D 

PROVIDER: S-EPMC7663484 | biostudies-literature | 2020 Oct

REPOSITORIES: biostudies-literature