Core Technician: Sierra Anderson
Polarized epithelial cell lines maintained and available: human intestinal T84, HT29, Caco2-BBe, and SKCO-15; canine renal MDCK I and MDCK II cells; porcine kidney LL-CPK cells
Second site storage available for specialized cell lines prepared by HDDC members:
This involves expanding the cell line, testing for mycoplasma, and frozen storage (if clear of mycoplasma) in duplicate vials in two separate LN2 dewars nCounter NanoString RNA Analysis
- Special polarized epithelial cell culture projects with discussion.
Epithelial cells for polarized culture are plated once a week based upon requests from individual investigators. Monolayers are then grown to full differentiation. Lentiviral and plasmid systems are available and the Core can assist in gene transduction of all cell lines. CRISPR technology is in place for effective gene editing of Caco-2 BBe and other cells. Genetic manipulations and culture services for selecting and expanding stable clones after transduction are provided upon approval by the Core Director, based on the availability of core personnel.
All cell lines are characterized by phenotype (morphology and electrical physiology) and proven mycoplasma free by PCR before and after storage. Actively passaged cell lines are tested for mycoplasma at least once every 3 months.