In Vitro Generation of Mouse Colon Crypts.

Organoid culture has had a significant impact on studies of the intestinal epithelium; however, the exquisite architecture, luminal accessibility, and lineage compartmentalization found has not been recapitulated in the organoid systems. We have used a microengineered platform with suitable extracellular matrix contacts and stiffness to generate a self-renewing mouse colonic epithelium that replicates key architectural and physiological functions found , including a surface lined with polarized crypts. Chemical gradients applied to the basal-luminal axis compartmentalized the stem/progenitor cells and promoted appropriate lineage differentiation along the crypt axis so that the tissue possessed a crypt stem cell niche as well as a layer of differentiated cells covering the luminal surface. This new approach combining microengineered scaffolds, native chemical gradients, and biophysical cues to control primary epithelium can serve as a highly functional and physiologically relevant tissue model.