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.
Footnotes
The authors declare no competing financial interests.
This work was supported by National Institutes of Health Grants R01CA196885 (to G.M.), R21NS094809 (to K.E.Z.), R01NS098772 and R01DA042852 (to R.K.), and R01NS095994 (to A.A.K.); the Children’s Tumor Foundation NF1 Synodos Grant 2015-04-009A (to R.K.); the National Cancer Institute Cancer Center Support Grant P30CA023074 Experimental Mouse Resource Service; and BIO5 Institute, University of Arizona. S.P. and this research was supported by a Science Foundation AZ Bisgrove Scholars postdoctoral fellowship.
This is an open-access article distributed under the terms of the Creative Commons Attribution 4.0 International license, which permits unrestricted use, distribution and reproduction in any medium provided that the original work is properly attributed.
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