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Simplified Murine 3D Neuronal Cultures for Investigating Neuronal Activity and Neurodegeneration

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Abstract

The ability to model brain tissue in three-dimensions offers new potential for elucidating functional cellular interactions and corruption of such functions during pathogenesis. Many protocols now exist for growing neurones in three-dimensions and these vary in complexity and cost. Herein, we describe a straight-forward method for generating three-dimensional, terminally differentiated central nervous system cultures from adult murine neural stem cells. The protocol requires no specialist equipment, is not labour intensive or expensive and produces mature cultures within 10 days that can survive beyond a month. Populations of functional glutamatergic neurones could be identified within cultures. Additionally, the three dimensional neuronal cultures can be used to investigate tissue changes during the development of neurodegenerative disease where demonstration of hallmark features, such as plaque generation, has not previously been possible using two-dimensional cultures of neuronal cells. Using a prion model of acquired neurodegenerative disease, biochemical changes indicative of prion pathology were induced within 2–3 weeks in the three dimensional cultures. Our findings show that tissue differentiated in this simplified three dimensional culture model is physiologically competent to model central nervous system cellular behaviour as well as manifest the functional failures and pathological changes associated with neurodegenerative disease.

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Acknowledgments

This work was funded by an NHMRC programme grant (SJC, #628946) and an NHMRC project grant (CLH, #APP1044264). SJC is supported by an NHMRC Practitioner Fellowship (#APP100581) and CLH is supported by a CJDSGN Memorial Grant in memory of Rhonda McCoy and the many people affected by prion disease in Australia.

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Authors and Affiliations

  1. Department of Medicine (Royal Melbourne Hospital), Melbourne Brain Centre, The University of Melbourne, 30 Royal Parade, Parkville, Melbourne, VIC, 3010, Australia

    Steven J. Collins & Cathryn L. Haigh

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  1. Steven J. Collins
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  2. Cathryn L. Haigh
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Correspondence to Cathryn L. Haigh.

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Compliance with Ethical Standards

NSCs were harvested as approved by the University of Melbourne animal ethics committee (ID 1011746), in compliance with Australian code of practice for the care and use of animals for scientific purposes.

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The authors declare that they have no competing interests.

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Collins, S.J., Haigh, C.L. Simplified Murine 3D Neuronal Cultures for Investigating Neuronal Activity and Neurodegeneration. Cell Biochem Biophys 75, 3–13 (2017). https://doi.org/10.1007/s12013-016-0768-z

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  • DOI: https://doi.org/10.1007/s12013-016-0768-z

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