Long-term expansion of epithelial organoids from human colon, adenoma, adenocarcinoma, and Barrett's epithelium

TY - JOUR

T1 - Long-term expansion of epithelial organoids from human colon, adenoma, adenocarcinoma, and Barrett's epithelium

AU - Sato, T.

AU - Stange, D.E.

AU - Ferrante, M.

AU - Vries, R.G.J.

AU - van Es, J.H.

AU - van den Brink, S.

AU - Houdt, W.J.

AU - Pronk, A.

AU - van Gorp, J.

AU - Siersema, P.D.

AU - Clevers, H.

N1 - Reporting year: 2011

PY - 2011

Y1 - 2011

N2 - BACKGROUND & AIMS: We previously established long-term culture conditions under which single crypts or stem cells derived from mouse small intestine expand over long periods. The expanding crypts undergo multiple crypt fission events, simultaneously generating villus-like epithelial domains that contain all differentiated types of cells. We have adapted the culture conditions to grow similar epithelial organoids from mouse colon and human small intestine and colon. METHODS: Based on the mouse small intestinal culture system, we optimized the mouse and human colon culture systems. RESULTS: Addition of Wnt3A to the combination of growth factors applied to mouse colon crypts allowed them to expand indefinitely. Addition of nicotinamide, along with a small molecule inhibitor of Alk and an inhibitor of p38, were required for long-term culture of human small intestine and colon tissues. The culture system also allowed growth of mouse Apc-deficient adenomas, human colorectal cancer cells, and human metaplastic epithelia from regions of Barrett's esophagus. CONCLUSIONS: We developed a technology that can be used to study infected, inflammatory, or neoplastic tissues from the human gastrointestinal tract. These tools might have applications in regenerative biology through ex vivo expansion of the intestinal epithelia. Studies of these cultures indicate that there is no inherent restriction in the replicative potential of adult stem cells (or a Hayflick limit) ex vivo.

AB - BACKGROUND & AIMS: We previously established long-term culture conditions under which single crypts or stem cells derived from mouse small intestine expand over long periods. The expanding crypts undergo multiple crypt fission events, simultaneously generating villus-like epithelial domains that contain all differentiated types of cells. We have adapted the culture conditions to grow similar epithelial organoids from mouse colon and human small intestine and colon. METHODS: Based on the mouse small intestinal culture system, we optimized the mouse and human colon culture systems. RESULTS: Addition of Wnt3A to the combination of growth factors applied to mouse colon crypts allowed them to expand indefinitely. Addition of nicotinamide, along with a small molecule inhibitor of Alk and an inhibitor of p38, were required for long-term culture of human small intestine and colon tissues. The culture system also allowed growth of mouse Apc-deficient adenomas, human colorectal cancer cells, and human metaplastic epithelia from regions of Barrett's esophagus. CONCLUSIONS: We developed a technology that can be used to study infected, inflammatory, or neoplastic tissues from the human gastrointestinal tract. These tools might have applications in regenerative biology through ex vivo expansion of the intestinal epithelia. Studies of these cultures indicate that there is no inherent restriction in the replicative potential of adult stem cells (or a Hayflick limit) ex vivo.

U2 - 10.1053/j.gastro.2011.07.050

DO - 10.1053/j.gastro.2011.07.050

M3 - Article

SN - 0016-5085

VL - 141

SP - 1762

EP - 1772

JO - Gastroenterology

JF - Gastroenterology

IS - 5

ER -