From Cell Culture to Organoids-Model Systems for Investigating Prion Strain Characteristics

Pineau, Hailey; Sim, Valerie L.

doi:10.3390/biom11010106

“…Refinement, reduction and replacement (“3R”) of animal studies is desirable due to ethical reasons but also towards less cost and labour-intensive methods, therefore alternatives to bioassays such as versatile cell culture models are needed. Already early in prion research a first persistently infected cell line from mouse was established 7 , later further cell models were introduced (for review see 8 ). 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 .…”

Section: Introductionmentioning

confidence: 99%

“…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 . However, 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 only 8 , 10 .…”

Section: Introductionmentioning

confidence: 99%

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

Schwenke

¹

,

Wälzlein

²

,

Bauer

³

et al. 2022

Sci Rep

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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.

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“…To verify our findings, we established RK13 cell lines that stably expressed WT, ΔN6, or Met4-1 PrPs. RK13 is a rabbit kidney epithelial cell line that expresses little endogenous PrP and has been widely used to determine the susceptibility of various PrP forms to prion infection ( 36 , 37 ).…”

Section: Resultsmentioning

confidence: 99%

“…Since N2a neuroblastoma cell line is susceptible to prion infection (37), we used it as a positive control and found that the cellular localization of transfected PrPs were similar to J o u r n a l P r e -p r o o f that of endogenous PrP in N2a cells (Supporting Fig. S1).…”

Section: Expression Of Wt Met4-1 and ∆N6 Prps In Rk13 Cellsmentioning

confidence: 99%

The protease-sensitive N-terminal polybasic region of prion protein modulates its conversion to the pathogenic prion conformer

Zhang

¹

,

Pan

²

,

Ying

³

et al. 2021

Journal of Biological Chemistry

2

0

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“…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]. However, 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 only [10,8].…”

Section: Introductionmentioning

confidence: 99%

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

Schwenke

¹

,

Waelzlein

²

,

Bauer

³

et al. 2021

Preprint

1

0

View full text Add to dashboard Cite

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 in a single cell assay.

show abstract

From Cell Culture to Organoids-Model Systems for Investigating Prion Strain Characteristics

Cited by 10 publications

References 131 publications

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

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

The protease-sensitive N-terminal polybasic region of prion protein modulates its conversion to the pathogenic prion conformer

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

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