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eNeuro. 2018 Sep 27;5(5). pii: ENEURO.0135-18.2018. doi: 10.1523/ENEURO.0135-18.2018. eCollection 2018 Sep-Oct.

High Fidelity Cryopreservation and Recovery of Primary Rodent Cortical Neurons.

Author information

1
Department of Neuroscience, College of Science, University of Arizona, Tucson, AZ 85724.
2
Department of Cellular and Molecular Medicine, College of Medicine, University of Arizona, Tucson, AZ 85724.
3
Department of Pharmacology, College of Medicine, University of Arizona, Tucson, AZ 85724.
4
Bio5 Institute, University of Arizona, Tucson, AZ 85724.
5
Department of Molecular and Cellular Biology, College of Science, University of Arizona, Tucson, AZ 85724.
6
Departments of Neurology, Cellular and Molecular Medicine, and Immunobiology, College of Medicine, University of Arizona, Tucson, AZ 85724.
7
Departments of Neuroscience, Pharmacology, and Anesthesiology, Colleges of Science and Medicine, University of Arizona, Tucson, AZ 85724.
8
Departments of Neuroscience and Molecular and Cellular Biology, College of Science, University of Arizona, Tucson, AZ 85724.

Abstract

Cell cryopreservation improves reproducibility and enables flexibility in experimental design. Although conventional freezing methodologies have been used to preserve primary neurons, poor cell viability and reduced survival severely limited their utility. We screened several high-performance freezing media and found that CryoStor10 (CS10) provided superior cryoprotection to primary mouse embryonic cortical neurons compared to other commercially-available or traditional reagents, permitting the recovery of 68.8% of cells relative to a fresh dissection. We characterized developmental, morphometric, and functional indicators of neuron maturation and found that, without exception, neurons recovered from cryostorage in CS10 media faithfully recapitulate in vitro neurodevelopment in-step with neurons obtained by fresh dissection. Our method establishes cryopreserved neurons as a reliable, efficient, and equivalent model to fresh neuron cultures.

KEYWORDS:

cryopreservation; primary neuron culture

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