The human rectum is present in the lower gastrointestinal tract and functions as a temporary storage site for fecal material. Rectal fibroblasts are important structural components of the rectum and involve a number of diseases; such as ulcerative colitis, Crohn’s disease, and rectal cancer. The main functions of fibroblasts are to maintain the structural integrity of connective tissue and also to aid in tissue repair and remodeling. Functional changes to human rectal fibroblasts (HRecF) can impair wound healing and affect epithelial cell proliferation in ulcerative colitis patients. In Crohn’s disease, HRecF respond to intestinal inflammation by proliferating abnormally, which leads to fibrotic scarring. Primary HRecF are useful tools to study the pathophysiology of several disesases; such as rectal cancer, ulcerative colitis, and Crohn’s disease and to help develop therapies for these diseases.