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

Details

Serval ID
serval:BIB_BE43928C941E
Type
Article: article from journal or magazin.
Collection
Publications
Institution
Title
Enzymatic Dissociation Induces Transcriptional and Proteotype Bias in Brain Cell Populations.
Journal
International journal of molecular sciences
Author(s)
Mattei D., Ivanov A., van Oostrum M., Pantelyushin S., Richetto J., Mueller F., Beffinger M., Schellhammer L., Vom Berg J., Wollscheid B., Beule D., Paolicelli R.C., Meyer U.
ISSN
1422-0067 (Electronic)
ISSN-L
1422-0067
Publication state
Published
Issued date
26/10/2020
Peer-reviewed
Oui
Volume
21
Number
21
Pages
E7944
Language
english
Notes
Publication types: Journal Article
Publication Status: epublish
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
astrocytes, enzymatic digestion, microglia, microglia isolation, neurons, protocol, single-cell sequencing
Pubmed
Open Access
Yes
Create date
09/11/2020 13:29
Last modification date
13/11/2020 6:26
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