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Journal of Virology, June 2000, p. 5597-5603, Vol. 74, No. 12
Section of Infectious Diseases, The Children's Hospital of
Philadelphia,1 The University of
Pennsylvania School of Medicine,2
The University of Pennsylvania School of Veterinary
Medicine,4 and The Wistar Institute of
Anatomy and Biology,5 Philadelphia,
Pennsylvania 19104, and Georgetown University School of
Medicine, Washington, D.C. 200073
Received 9 December 1999/Accepted 13 March 2000
We describe a method for long-term culture of primary small
intestinal epithelial cells (IEC) from suckling mice. IEC were digested
from intestinal fragments as small intact units of epithelium (organoids) by using collagenase and dispase. IEC proliferated from
organoids on a basement-membrane-coated culture surface and remained
viable for 3 weeks. Cultured IEC had the morphologic and functional
characteristics of immature enterocytes, notably sustained expression
of cytokeratin and alkaline phosphatase. Few mesenchymal cells were
present in the IEC cultures. IEC were also cultured from adult BALB/c
mice and expressed major histocompatibility complex (MHC) class II
antigens for at least 48 h in vitro. Primary IEC supported the
growth of rhesus rotavirus (RRV) to a greater extent than a murine
small intestinal cell line, m-ICcl2. Cell-culture-adapted murine rotavirus strain EDIM infected primary IEC and
m-ICcl2 cells to a lesser extent than RRV. Wild-type EDIM
did not infect either cell type. Long-term culture of primary murine
small intestinal epithelial cells provides a method to study (i)
virus-cell interactions, (ii) the capacity of IEC to act as
antigen-presenting cells using a wide variety of MHC haplotypes, and
(iii) IEC biology.
0022-538X/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.
Primary Murine Small Intestinal Epithelial Cells,
Maintained in Long-Term Culture, Are Susceptible to Rotavirus
Infection
*
Corresponding author. Mailing address: Division of
Immunologic and Infectious Diseases, The Children's Hospital of
Philadelphia, Abramson Research Bldg., 12th Floor, 34th St. and Civic
Center Blvd., Philadelphia, PA 19104. Phone: (215) 590-2186. Fax:
(215) 590-2025. E-mail: macartney{at}emailchop.edu.
Journal of Virology, June 2000, p. 5597-5603, Vol. 74, No. 12
0022-538X/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.
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