Title:
Development of human intestinal stem cell culture system
Toshiro Sato MD, PhD
Division of Gastroenterology and Hepatology,
Department of Internal Medicine, Keio University School of Medicine
Abstract:
The intestinal epithelium is the most rapidly self-renewing tissue in adult mammals. Lgr5+ stem cells are located at crypt bottom and identified as intestinal stem cells. We developed a new culture system in which intestinal stem cells form organoid structure showing stem cell self-renewal and multiple lineages differentiation in the absence of non-intestinal epithelial cells. We proved that Paneth cells, one of the daughter cells from intestinal stem cells, produce essential growth factors (Wnt-3, EGF and Notch ligand) for intestinal stem cells and function as intestinal stem cell niche.
Recently, we developed new organoid culture systems for other digestive organs (stomach, pancreas and liver) and human tissues. Each organoid culture requires basic growth factors mixture (EGF, noggin and R-spondin) and organ specific growth factors (e.g. HGF for liver). The culture technology should be widely applicable as a research tool for infectious, inflammatory and neoplastic pathologies of the human gastrointestinal tract. Moreover, regenerative applications may become feasible with ex vivo expanded intestinal epithelia.
References:
1) Jung P, Sato T, TM, Merlos-Suarez A, Barriga FM, Iglesias M, Gallardo MM, Blasco MA, Sancho E, Clevers H, Batlle E. Isolation and in vitro expansion of human colonic stem cells. Nature Medicine 2011 ； 17:1225-7.
2) Sato T, Stange DE, Vries RGJ, van ES JH, van den Brink S, van Houdt WJ, Pronk A, van Gorp J, Siersema PD, Clevers H ． Long-term in vitro expansion of epithelial organoids from human colon, adenoma, adenocarcinoma and Barrett ‘ s Esophagus. Gastroenterology.2011;141:1762-72.
3) Sato T, van Es JH, Snippert HJ, Stange DE, Vries RG, van den Born M, Barker N, Shroyer NF, van de Wetering M, Clevers H. Paneth cells constitute the niche for Lgr5 stem cells in intestinal crypts. Nature. 2011;469:415-8.
4) Sato T , Vries RG, Snippert HJ, van de Wetering M, Barker N, Stange DE, van Es JH, Abo A, Kujala P, Peters PJ, Clevers H. Single Lgr5 stem cells build crypt-villus structures in vitro without a mesenchymal niche. Nature . 2009;459:262-5.
