Establishing Mixed Neuronal and Glial Cell Cultures from Embryonic Mouse Brains to Study Infection and Innate Immunity

The ultimate research question of my project is to figure out how we can manipulate the innate immune responses of the central nervous system to protect against viral infections. Currently, we research their response to the profoundly antiviral protein interferon beta. An experimental challenge is generating enough cells to allow for detailed investigation.

Each result needs to be validated using multiple techniques and this can, unfortunately, result in many animals being culled. Due to the large number of cells generated by this novel culture technique, many different experimental conditions can be investigated from one pregnant mouse. Additionally, using in vitro alternatives, where possible, also greatly decreases animal suffering.

This method reduces and refines animal use, closely following the principles of the three Rs.This culture method can be used to address many different research questions using a variety of readout methods such as ELISA, qPCR, microscopy and flow cytometry. This technique could help other researchers to reduce and refine their animal usage. We hope to improve our understanding of the innate antiviral response, modulating it to protect against opportunistic viruses such as human polyomavirus 2, or John Cunningham virus, as it is commonly known, the primary agent of PML which can be a severe side effect of immune suppressive drugs used to treat MS and other diseases.