Intestinal Enteroid Culture for Human Astroviruses

Irene A Owusu; Carmen Mirabelli; Abimbola O Kolawole

doi:10.21769/BioProtoc.3687

Intestinal Enteroid Culture for Human Astroviruses

Bio Protoc. 2020 Jul 20;10(14):e3687. doi: 10.21769/BioProtoc.3687.

Authors

Irene A Owusu^{1

2}, Carmen Mirabelli¹, Abimbola O Kolawole³

Affiliations

¹ Department of Microbiology and Immunology, University of Michigan, Ann Arbor, Michigan, USA.
² West African Center for Cell Biology of Infectious Pathogens, University of Ghana, Legon, Ghana.
³ Department of Biological Sciences, Wright State University, Dayton, Ohio, USA.

Abstract

Human astroviruses (HAstV) are non-enveloped, positive-sense single stranded RNA viruses that typically cause gastroenteritis in children, the elderly and among immunocompromised individuals. Some HAstV species have also been implicated in neurological diseases. It is important to study these viruses to understand the pathogenesis and develop therapeutics. Here we describe HAstV infection in epithelium-only human intestinal enteroids (HIE) isolated from biopsy-derived intestinal crypts. Although different HAstV clades have been propagated in transformed immortalized cell lines such as A549, Caco-2, HEK293T and Huh7.5, we chose HIE because they better mimic the human intestine and thus are more physiologically relevant. Additionally, HIE support the replication of all HAstV clades including clinical samples, thus making HIE a valuable potential universal model to study HAstV biology.

Keywords: Astrovirus; Enteric virus; Enteroid; HAstV; Infection; MLB; VA.

Grants and funding

R21 AI141835/AI/NIAID NIH HHS/United States