A Strainer-Based Platform for the Collection and Immunolabeling of Porcine Epidemic Diarrhea Virus-Infected Porcine Intestinal Organoid

Int J Mol Sci. 2023 Oct 27;24(21):15671. doi: 10.3390/ijms242115671.

Abstract

The development of organoid research has raised new requirements for this methodology. In a previous study, we demonstrated that an emerging protocol achieved the collection, loading, and programmed immunolabeling of mouse intestinal organoids based on a strainer platform. To uncover the applied potential of this novel methodology on organoids from other species, the strainer platform was utilized to characterize the porcine epidemic diarrhea virus (PEDV)-infected porcine intestinal organoid model. Based on a previous study, some steps were changed to improve the efficiency of the assay by simplifying the reagent addition procedure. In addition, we redefined the range of strainer sizes on porcine intestinal organoids, showing that strainers with pore sizes of 40 and 70 μm matched the above protocol well. Notably, the strainer platform was successfully used to label viral proteins, laying the foundation for its application in the visualization of viral infection models. In summary, the potential of the strainer platform for organoid technology was explored further. More extensive exploration of this platform will contribute to the development of organoid technology.

Keywords: collection; labeling; porcine epidemic diarrhea virus; porcine intestinal organoid; strainer platform.

MeSH terms

  • Animals
  • Coronavirus Infections*
  • Diarrhea
  • Intestines
  • Mice
  • Organoids
  • Porcine epidemic diarrhea virus*
  • Swine
  • Swine Diseases*
  • Viral Proteins

Substances

  • Viral Proteins