Culture medium for colonic organoids differs from that for small intestinal organoids (e.g. Wnt3a growth factor). Does your medium contain these extra growth factors, or do they need to be added separately? Also, does the described isolation protocol differ between small intestinal and colon organoids?

Answer

Indeed the culture of colon crypts differs from that of the small intestine. We have found that IntestiCult™ Organoid Growth Medium (Mouse) works on both tissues. In the primary culture, the efficiency of cultured colon crypts that grow into mature organoids may not be as high as the efficiency of the small intestinal cultures, but it does increase upon passaging. The described isolation protocol works well with the exception of substituting the dissection of the colon rather than small intestine.