Colonic myofibroblast cell line stimulates colonoid formation

Abstract

A stable and efficient system for the culture of murine colon epithelial cells or crypts is required to facilitate studies of the dynamics and factors affecting colon stem cell niche and crypt formation. Survival of colonic epithelial cells or crypts in vitro was not established until recently, when it was found that exogenous Wnt3A and R-spondin could promote cell survival and formation of spheroids (colonospheres) or some advanced organoids with well-developed crypts (colonoids). However, after 6-8 days in these culture conditions, only small numbers of colonospheres form organoids with crypt-like structures (colonoids). This study describes the use of a myofibroblast cell line and a coculture system that increases the efficiency of colonoid formation from isolated crypts. The enhanced coculture system has significantly improved colonoid-forming efficiency compared with results from previous systems. Crypt formation can be detected as early as day 2. The coculture system will facilitate the characterization of the colon stem cell niche and the changes that occur as a result of perturbations or mutations in colon stem or epithelial cells, such as those that favor precancerous adenoma or cancer.

Retinol Promotes In Vitro Growth of Proximal Colon Organoids through a Retinoic Acid-Independent Mechanism

“…This methodological advance has led to new approaches to investigate the mechanism of proliferation of normal colonic epithelial cells [37–39] and also the pathways of their tumorigenesis [40–42]. To our knowledge, however, there has been no study that assessed ex vivo response of colonic epithelial stem cell populations of different parts along the length of the colon and described their different behaviors.…”

Analysis of Wnt signalling dynamics during colon crypt development in 3D culture

“…1C). Single crypts are not observed in the conventional 3D cultures where the crypts or fragments are embedded in Matrigel2123.…”

Tissue Engineering Functional Gastrointestinal Regions: The Importance of Stem and Progenitor Cells

“…Coculture with myofibroblasts has been shown to improve colonocyte growth (Hirokawa et al 2014). Similar to BMP suppression of the Wnt pathway, the TGF-b suppression of Wnt inhibits colonic crypt proliferation, whereas TGF-b inhibition with A83-01 stimulates growth (Reynolds et al 2014).…”