Enzymatic dissociation induces transcriptional and proteotype bias in brain cell populations
Item Type: | Preprint |
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Title: | Enzymatic dissociation induces transcriptional and proteotype bias in brain cell populations |
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Creators Name: | Mattei, D. and Ivanov, A. and van Oostrum, M. and Pantelyushin, S. and Richetto, J. and Mueller, F. and Beffinger, M. M. and Schellhammer, L. and vom Berg, J. and Wollscheid, B. and Beule, D. and Paolicelli, R.C. and Meyer, U. |
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Abstract: | Different cell isolation techniques exist for transcriptomic and proteotype profiling of brain cells. Here, we provide asystematic 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 artefacts when implementing enzymatic digestion-based isolation methods for brain cell analyses. |
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Keywords: | Microglia, Astrocytes, Transcriptomics, Proteotype, Brain Tissue, Dissociation, Animals, Mice |
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Source: | bioRxiv |
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Title of Book: | Enzymatic dissociation induces transcriptional and proteotype bias in brain cell populations |
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Publisher: | Cold Spring Harbor Laboratory Press |
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Article Number: | 2020.05.14.095422 |
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Date: | 16 May 2020 |
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Official Publication: | https://doi.org/10.1101/2020.05.14.095422 |
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