Proteomics

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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 show that standard enzymatic digestion of brain tissue at 37°C induces profound alterations in the transcriptome and proteotype of specific brain cells, as compared to cold mechanical dissociation. These findings emphasize the need to consider intrinsic bias and biological artefacts when implementing enzymatic digestion-based isolation methods for brain cell analyses.

INSTRUMENT(S): Orbitrap Fusion Lumos, Orbitrap Fusion

ORGANISM(S): Mus Musculus (mouse)

TISSUE(S): Brain

SUBMITTER: Marc van Oostrum  

LAB HEAD: Bernd Wollscheid

PROVIDER: PXD015592 | Pride | 2021-09-08

REPOSITORIES: Pride

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

Mattei Daniele D   Ivanov Andranik A   van Oostrum Marc M   Pantelyushin Stanislav S   Richetto Juliet J   Mueller Flavia F   Beffinger Michal M   Schellhammer Linda L   Vom Berg Johannes J   Wollscheid Bernd B   Beule Dieter D   Paolicelli Rosa Chiara RC   Meyer Urs U  

International journal of molecular sciences 20201026 21

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  ...[more]

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