Fibroblasts enhance the growth and survival of adult feline small intestinal organoids

Abstract Intestinal organoids are important cell culture models that complement live animal studies of intestinal biology. Adult feline small intestinal organoids are needed for infectious disease research but are difficult to work with due to slow growth and rapid senescence. We introduce a method of co-culturing adult feline small intestinal organoids with growth inhibited human foreskin fibroblast feeder cells to enhance organoid proliferation and survival. With feeder cells, feline jejunal and ileal organoids survived at least nine months in culture until cryopreservation. Fibroblast supplementation also increased the maximum size of cat and mouse intestinal organoids, making them easier for microinjection. The increased proliferation, longevity, and size make fibroblast-supplementation of feline small intestinal organoids a significant improvement on current methods. These methods have high potential to reduce the number of cats used for research and may be applicable for intestinal cells from other animals that are difficult to culture.