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| Item Type: | Article |
|---|---|
| Title: | Enzymatic dissociation induces transcriptional and proteotype bias in brain cell populations |
| Creators: |
Mattei, D.  ORCID: https://orcid.org/0000-0001-6221-9321, Ivanov, A. ORCID: https://orcid.org/0000-0002-4824-1925, van Oostrum, M. ORCID: https://orcid.org/0000-0001-8747-9787, Pantelyushin, S. ORCID: https://orcid.org/0000-0002-3490-3860, Richetto, J. ORCID: https://orcid.org/0000-0002-8313-3348, Mueller, F., Beffinger, M., Schellhammer, L., Vom Berg, J. ORCID: https://orcid.org/0000-0002-4086-1564, Wollscheid, B. ORCID: https://orcid.org/0000-0002-3923-1610, Beule, D. ORCID: https://orcid.org/0000-0002-3284-0632, Paolicelli, R.C. ORCID: https://orcid.org/0000-0001-6851-0849 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: | 21 |
| Number: | 21 |
| Page Range: | 7944 |
| Date: | 1 November 2020 |
| Official Publication: | https://doi.org/10.3390/ijms21217944 |
| PubMed: | View item in PubMed |
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