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Methanol fixation is the method of choice for droplet-based single-cell transcriptomics of neural cells

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The main critical step in single-cell transcriptomics is sample preparation. Several methods have been developed to preserve cells after dissociation to uncouple sample handling from library preparation. Yet, the suitability of these methods depends on the cell types to be processed. In this project, we perform a systematic comparison of preservation methods for droplet-based single-cell RNA-seq on neural and glial cells derived from induced pluripotent stem cells. Our results show that while DMSO provides the highest cell quality in terms of RNA molecules and genes detected per cell, it strongly affects the cellular composition and induces the expression of stress and apoptosis genes. In contrast, methanol fixed samples display a cellular composition similar to fresh samples and provide a good cell quality and little expression biases. Taken together, our results show that methanol fixation is the method of choice for performing droplet-based single-cell transcriptomics experiments on neural cell populations. Methanol fixation appears to be the method of choice for droplet-based scRNA-seq on neural cell populations, based on a broad analysis of how various fixation or preservation methods impact the single-cell transcriptomes of hiPSC-derived neural cells.

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Communications Biology, 2023, vol. 6, num. 1

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cc by (c) Gutiérrez Franco, Ana et al, 2023

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Articles publicats en revistes (Institut d'lnvestigació Biomèdica de Bellvitge (IDIBELL))

# metadata.dc.title[].value
Methanol fixation is the method of choice for droplet-based single-cell transcriptomics of neural cells