Enzymatic dissociation induces transcriptional and proteotype bias in brain cell populations

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Item Type:

Article

Title:

Enzymatic dissociation induces transcriptional and proteotype bias in brain cell populations

Creators Name:

Mattei, D. and Ivanov, A. and van Oostrum, M. and Pantelyushin, S. and Richetto, J. and Mueller, F. and Beffinger, M. and Schellhammer, L. and Vom Berg, J. and Wollscheid, B. and Beule, D. and Paolicelli, R.C. and Meyer, U.

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 isolation methods for brain cell analyses.

Keywords:

Microglia, Astrocytes, Neurons, Enzymatic Digestion, Single-Cell Sequencing, Protocol, Microglia Isolation, Animals, Mice

Source:

International Journal of Molecular Sciences

ISSN:

1422-0067

Publisher:

MDPI

Volume:

Number:

Page Range:

7944

Date:

1 November 2020

Official Publication:

https://doi.org/10.3390/ijms21217944

PubMed:

View item in PubMed

Related to:

URL	URL Type
https://edoc.mdc-berlin.de/18948/	Preprint version

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