Spontaneous maturation of human colonic organoids (1119.2)

The intestinal epithelium is a complex structure consisting of distinct cell types derived from stem cells residing in the crypt base. Here we investigated the spontaneous maturation of organoids grown from human colonic crypts. Crypts were isolated by Ca2+ chelation from proximal colonic biopsies and transferred to Matrigel and growth media. The organoids were grown for 15 days and the gross appearance, ultrastructure, histology of the organoids and gene expression of proliferation, differentiation, maturation and stem cell activity markers analysed at day 4 and 15 by qPCR. At day 4, 71±7% of the organoids were thin walled spheroids, while at day 15 there was a mixture of large convoluted organoids (51±10%) and spheroids. At day 4 the organoids consisted of thin undifferentiated cells, whereas at day 15 the convoluted organoids consisted of differentiated columnar epithelial cells. Associated with this there was an increase (p < 0.0001) in expression of mature enterocyte and goblet cell differentiation m...