Primary murine small intestinal epithelial cells, maintained in long-term culture, are susceptible to rotavirus infection

We describe a method for long-term culture of primary small intestinal epit helial cells (IEC) from suckling mice. IEC were digested from intestinal fr agments as small intact units of epithelium (organoids) by using collagenas e and dispase. IEC proliferated from organoids on a basement-membrane coate d culture surface and remained viable for 3 weeks. Cultured IEC had the mor phologic and functional characteristics of immature enterocytes, notably su stained 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 a ntigens for at least 48 h in vitro. Primary IEC supported the growth of rhe sus rotavirus (RRV) to a greater extent than a murine small intestinal cell line, m-ICc12 Cell-culture-adapted murine rotavirus strain EDIM infected p rimary IEC and m-ICc12 cells to a lesser extent than RRV. Wild-type EDIM di d not infect either cell type. Long-term culture of primary murine small in testinal epithelial cells provides a method to study (i) virus-cell interac tions, (ii) the capacity of IEC to act as antigen-presenting cells using a wide variety of MHC haplotypes, and (iii) IEC biology.