Primary glia cells from bank vole propagate multiple rodent-adapted scrapie prions

Karla A Schwenke,Achim Thomzig,Agnieszka Bauer,Michael Beekes,Joo-Hee Wälzlein

doi:10.1038/s41598-022-06198-4

Abstract

Since the beginning prion research has been largely dependent on animal models for deciphering the disease, drug development or prion detection and quantification. Thereby, ethical as well as cost and labour-saving aspects call for alternatives in vitro. Cell models can replace or at least complement animal studies, but their number is still limited and the application usually restricted to certain strains and host species due to often strong transmission barriers. Bank voles promise to be an exception as they or materials prepared from them are uniquely susceptible to prions from various species in vivo, in vitro and in cell-free applications. Here we present a mainly astrocyte-based primary glia cell assay from bank vole, which is infectible with scrapie strains from bank vole, mouse and hamster. Stable propagation of bank vole-adapted RML, murine 22L and RML, and hamster 263K scrapie is detectable from 20 or 30 days post exposure onwards. Thereby, the infected bank vole glia cells show similar or even faster prion propagation than likewise infected glia cells of the corresponding murine or hamster hosts. We propose that our bank vole glia cell assay could be a versatile tool for studying and comparing multiple prion strains with different species backgrounds combined in one cell assay.

Highlights

Since the beginning prion research has been largely dependent on animal models for deciphering the disease, drug development or prion detection and quantification
To the best of our knowledge this is the first report of a primary glia cell culture from bank vole, which shows a stable propagation of rodent-adapted scrapie prions of murine, hamster and bank vole origin
The primary mixed glia cell culture revealed a high content of astrocytic cells in flow cytometry, of which 94% belonged to the doublepositive cell population for the specific astrocyte markers glial fibrillary acidic protein (GFAP) and glutamate transporter 1 (GLT-1)

Summary

Introduction

Since the beginning prion research has been largely dependent on animal models for deciphering the disease, drug development or prion detection and quantification. We present a mainly astrocyte-based primary glia cell assay from bank vole, which is infectible with scrapie strains from bank vole, mouse and hamster. One successful example of a bioassay alternative is the standard scrapie cell assay based on subcloned neuroblastoma N2a cells, which was developed for titre determination of murine scrapie (22L and RML) reaching equal sensitivity while being significantly faster and cheaper[9] This assay only works for mouse-adapted scrapie strains, which is a common problem of most cell culture models: being limited to certain host-adapted prions o nly[8,10]. A primary bank vole cell assay infectible with prions could provide a tool for investigating a variety of prion strains from different host species under more natural conditions in a single cell model. We present a primary mainly astrocyte-based cell assay from bank vole, which shows stable propagation of four scrapie strains from bank vole, mouse and hamster

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Conclusion