Related Experiment Video
Updated: Dec 17, 2025
Author Spotlight: Insight Into Advances in Prion Diseases Research
Published on: August 11, 2023
An astrocyte cell line that differentially propagates murine prions
Waqas Tahir1,2,3, Basant Abdulrahman1,2,3,4, Dalia H Abdelaziz1,2,3,4
1Department of Comparative Biology & Experimental Medicine, Faculty of Veterinary Medicine, University of Calgary, Calgary, Alberta, Canada.
Abstract:
Prion diseases are fatal infectious neurodegenerative disorders in human and animals caused by misfolding of the cellular prion protein (PrPC) into the pathological isoform PrPSc Elucidating the molecular and cellular mechanisms underlying prion propagation may help to develop disease interventions. Cell culture systems for prion propagation have greatly advanced molecular insights into prion biology, but translation of in vitro to in vivo findings is often disappointing. A wider range of cell culture systems might help overcome these shortcomings. Here, we describe an immortalized mouse neuronal astrocyte cell line (C8D1A) that can be infected with murine prions. Both PrPC protein and mRNA levels in astrocytes were comparable with those in neuronal and non-neuronal cell lines permitting persistent prion infection. We challenged astrocytes with three mouse-adapted prion strains (22L, RML, and ME7) and cultured them for six passages. Immunoblotting results revealed that the astrocytes propagated 22L prions well over all six passages, whereas ME7 prions did not replicate, and RML prions replicated only very weakly after five passages. Immunofluorescence analysis indicated similar results for PrPSc Interestingly, when we used prion conversion activity as a readout in real-time quaking-induced conversion assays with RML-infected cell lysates, we observed a strong signal over all six passages, comparable with that for 22L-infected cells. These data indicate that the C8D1A cell line is permissive to prion infection. Moreover, the propagated prions differed in conversion and proteinase K-resistance levels in these astrocytes. We propose that the C8D1A cell line could be used to decipher prion strain biology.
Insights
Researchers developed a new mouse astrocyte cell line (C8D1A) for studying prion diseases. This model supports prion infection and propagation, offering a valuable tool for understanding prion strain diversity and developing interventions.
Area of Science:
- Neuroscience
- Infectious Diseases
- Cell Biology
Background:
- Prion diseases are fatal neurodegenerative disorders caused by misfolded prion proteins.
- Existing cell culture models have limitations in translating in vitro findings to in vivo scenarios.
- Developing diverse cell culture systems is crucial for advancing prion disease research.
Purpose of the Study:
- To introduce and characterize a novel immortalized mouse neuronal astrocyte cell line (C8D1A) for prion propagation studies.
- To assess the permissiveness of C8D1A cells to different mouse-adapted prion strains.
- To investigate the potential of C8D1A cells in deciphering prion strain biology.
Main Methods:
- Infection of C8D1A cells with three mouse-adapted prion strains (22L, RML, ME7).
- Culturing infected cells over six passages.
- Analysis of prion propagation using immunoblotting and immunofluorescence.
- Assessment of prion conversion activity via real-time quaking-induced conversion (RT-QuIC) assays.
Main Results:
- C8D1A cells demonstrated persistent prion infection and propagated 22L prions effectively over six passages.
- ME7 prions did not replicate, and RML prions showed weak replication in C8D1A cells.
- RT-QuIC assays revealed significant prion conversion activity in RML-infected C8D1A cells, comparable to 22L-infected cells.
- Propagated prions exhibited differences in conversion and proteinase K resistance.
Conclusions:
- The C8D1A cell line is permissive to prion infection and can be utilized for prion propagation.
- This cell line supports differential propagation of prion strains, highlighting its utility in studying prion strain diversity.
- C8D1A astrocytes offer a promising model for investigating the molecular mechanisms of prion strain biology and developing therapeutic strategies.
