Susceptibility of ovine bone marrow-derived mesenchymal stem cell spheroids to scrapie prion infection

Hernaiz, Adelaida (Universidad de Zaragoza) ; Cobeta, Paula ; Marín, Belén (Universidad de Zaragoza) ; Vázquez, Francisco José (Universidad de Zaragoza) ; Badiola, Juan José (Universidad de Zaragoza) ; Zaragoza, Pilar (Universidad de Zaragoza) ; Bolea, Rosa (Universidad de Zaragoza) ; Martín-Burriel, Inmaculada

Resumen: In neurodegenerative diseases, including prion diseases, cellular in vitro models appear as fundamental tools for the study of pathogenic mechanisms and potential therapeutic compounds. Two-dimensional (2D) monolayer cell culture systems are the most used cell-based assays, but these platforms are not able to reproduce the microenvironment of in vivo cells. This limitation can be surpassed using three-dimensional (3D) culture systems such as spheroids that more effectively mimic in vivo cell interactions. Herein, we evaluated the effect of scrapie prion infection in monolayer-cultured ovine bone marrow-derived mesenchymal stem cells (oBM-MSCs) and oBM-MSC-derived spheroids in growth and neurogenic conditions, analyzing their cell viability and their ability to maintain prion infection. An MTT assay was performed in oBM-MSCs and spheroids subjected to three conditions: inoculated with brain homogenate from scrapie-infected sheep, inoculated with brain homogenate from healthy sheep, and non-inoculated controls. The 3D conditions improved the cell viability in most cases, although in scrapie-infected spheroids in growth conditions, a decrease in cell viability was observed. The levels of pathological prion protein (PrPSc) in scrapie-infected oBM-MSCs and spheroids were measured by ELISA. In neurogenic conditions, monolayer cells and spheroids maintained the levels of PrPSc over time. In growth conditions, however, oBM-MSCs showed decreasing levels of PrPSc throughout time, whereas spheroids were able to maintain stable PrPSc levels. The presence of PrPSc in spheroids was also confirmed by immunocytochemistry. Altogether, these results show that a 3D culture microenvironment improves the permissiveness of oBM-MSCs to scrapie infection in growth conditions and maintains the infection ability in neurogenic conditions, making this model of potential use for prion studies.

Idioma: Inglés

DOI: 10.3390/ani13061043

Año: 2023

Publicado en: Animals 13, 6 (2023), 1043 [18 pp.]
ISSN: 2076-2615

Factor impacto JCR: 2.7 (2023)

Categ. JCR: VETERINARY SCIENCES rank: 16 / 167 = 0.096 (2023) - Q1 - T1
Categ. JCR: AGRICULTURE, DAIRY & ANIMAL SCIENCE rank: 10 / 80 = 0.125 (2023) - Q1 - T1

Factor impacto CITESCORE: 4.9 - Veterinary (all) (Q1) - Animal Science and Zoology (Q1)

Factor impacto SCIMAGO: 0.698 - Veterinary (miscellaneous) (Q1) - Animal Science and Zoology (Q1)

Financiación: info:eu-repo/grantAgreement/ES/DGA/A19-20R
Financiación: info:eu-repo/grantAgreement/ES/DGA-FSE/IIU-2023-2017
Financiación: info:eu-repo/grantAgreement/ES/MICINN/RTI2018-098711-B-I00

Tipo y forma: Artículo (Versión definitiva)

Área (Departamento): Área Genética (Dpto. Anatom.,Embri.Genét.Ani.)
Área (Departamento): Área Sanidad Animal (Dpto. Patología Animal)
Área (Departamento): Área Medicina y Cirugía Animal (Dpto. Patología Animal)