Author Spotlight: The 3D Culturing of Organoids from Murine Intestinal Crypts and a Single Stem Cell for Organoid Research

Our research focusing on stem cell biology, especially intestinal stem cell, present at the bottom of intestinal crypt. The aim is to answer how homeostasis is maintained by the stem cell in the intestine. This 3D organ culture derived from intestinal stem cell provides a powerful tool to study the proliferation, differentiation, and maintenance of stem cells.

Organ technology aids the culture of intestinal stem cell for a long time by retaining the self-renewal and differentiation potential. The organoid has been widely used for basic and translational research studies on intestinal fragility and past fragility. Our current challenge is to understand the coronary system involved in intestinal epithelial cell and stem cell.

Understanding biological processes triggered by objective cloning is of key importance. One of our significant findings is that signaling through cloning maintain the homeostasis of intestinal epithelial growth and differentiation. Our protocol describe a method for consistently isolating small intestinal crypts, and subsequent culture of 3D organoids to improve cryptal releasing rate.

We establish a mechanical isolation method involving vigorous shaking, after treatment with EDTA. EDTA and mechanical dissociation can be combined to improve cryptal yields. Additionally, proper skill can reduce virus contamination to a minimum, increasing the number of crypts.

Our findings suggest that the signaling through the muscular and organism receptors appear to work together to maintain the homeostasis of in intestinal epithelial growth and differentiation. This encourages us to hypothesize that the non-neurocoronary-arteritic system played an important role in the moderation of intestinal stem cell niche.