Enzymatic Dissociation Induces Transcriptional and Proteotype Bias in Brain Cell Populations.
Int J Mol Sci
; 21(21)2020 Oct 26.
Article
em En
| MEDLINE
| ID: mdl-33114694
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.
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MEDLINE
Assunto principal:
Neoplasias Encefálicas
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Astrócitos
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Microglia
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Perfilação da Expressão Gênica
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Enzimas
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Citometria de Fluxo
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Glioma
Idioma:
En
Ano de publicação:
2020
Tipo de documento:
Article