Long-term Expansion of Epithelial Organoids From Human Colon, Adenoma, Adenocarcinoma, and Barrett's Epithelium
Long-term Expansion of Epithelial Organoids From Human Colon, Adenoma, Adenocarcinoma, and Barrett's Epithelium
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DOI：
10.1053/j.gastro.2011.07.050
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发表时间：
2011-11-01
期刊：
影响因子：
29.4
通讯作者：
Clevers, Hans
中科院分区：
文献类型：
--
作者：
Sato, Toshiro;Stange, Daniel E.;Clevers, Hans
背景和目的：我们之前建立了长期培养条件，在该条件下，来自小鼠小肠的单个隐窝或干细胞可以长期扩增。扩张的隐窝经历多次隐窝裂变事件，同时产生包含所有分化类型细胞的绒毛样上皮结构域。我们调整了培养条件，以从小鼠结肠和人类小肠和结肠中培养类似的上皮类器官。方法：在小鼠小肠培养体系的基础上，优化小鼠和人结肠培养体系。结果：将 Wnt3A 添加到应用于小鼠结肠隐窝的生长因子组合中，使它们能够无限期扩张。人类小肠和结肠组织的长期培养需要添加烟酰胺以及 Alk 小分子抑制剂和 p38 抑制剂。该培养系统还允许小鼠 Apc 缺陷腺瘤、人类结直肠癌细胞和巴雷特食管区域的人类化生上皮细胞生长。结论：我们开发了一种技术，可用于研究人类胃肠道的感染、炎症或肿瘤组织。这些工具可能通过肠上皮细胞的离体扩张在再生生物学中得到应用。对这些培养物的研究表明，成体干细胞的离体复制潜力（或海弗利克极限）不存在固有的限制。
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.