Table 9:

MNV infection of intestinal enteroids

Problem	Possible cause	Solution
Poor recovery of enteroids from mice	Sub-optimal crypt dissociation. If cells are not treated long enough with collagenase, the crypts will not dissociate, and if cells are treated too long, cell viability will be poor.	Examine the cells under the microscope every 10 minutes during the 30 min dissociation period to ensure optimal dissociation. Adjust incubation time accordingly.
Inadequate monolayer on transwells	Transwell was not properly coated or plated with single-cell suspension	Pre-coat entire transwell surface evenly; pre-wet pipette tips, ensure cells are in a single cell suspension before adding to transwell; use lobular rather than spheroidal enteroids to create single cell suspension