Division of Pediatric General and Thoracic Surgery, Cincinnati Children's Hospital Medical Center, Cincinnati, OH 45229, USA.
Division of Developmental Biology, Cincinnati Children's Hospital Medical Center, Cincinnati, OH 45229, USA.
Development. 2023 May 1;150(9). doi: 10.1242/dev.201416. Epub 2023 May 5.
The in vitro differentiation of pluripotent stem cells into human intestinal organoids (HIOs) has served as a powerful means for creating complex three-dimensional intestinal structures. Owing to their diverse cell populations, transplantation into an animal host is supported with this system and allows the temporal formation of fully laminated structures, including crypt-villus architecture and smooth muscle layers that resemble native human intestine. Although the endpoint of HIO engraftment has been well described, here we aim to elucidate the developmental stages of HIO engraftment and establish whether it parallels fetal human intestinal development. We analyzed a time course of transplanted HIOs histologically at 2, 4, 6 and 8 weeks post-transplantation, and demonstrated that HIO maturation closely resembles key stages of fetal human intestinal development. We also utilized single-nuclear RNA sequencing to determine and track the emergence of distinct cell populations over time, and validated our transcriptomic data through in situ protein expression. These observations suggest that transplanted HIOs do indeed recapitulate early intestinal development, solidifying their value as a human intestinal model system.
多能干细胞体外分化为人类肠道类器官（HIOs）为构建复杂的三维肠道结构提供了有力手段。由于其具有多样化的细胞群体，该系统支持将其移植到动物宿主中，并允许在时间上形成完全分层的结构，包括隐窝-绒毛结构和类似于天然人肠道的平滑肌层。尽管 HIO 植入的终点已经得到很好的描述，但在这里，我们旨在阐明 HIO 植入的发育阶段，并确定其是否与胎儿人类肠道发育平行。我们在移植后 2、4、6 和 8 周对移植的 HIO 进行了组织学分析，结果表明 HIO 成熟非常类似于胎儿人类肠道发育的关键阶段。我们还利用单细胞 RNA 测序来确定和跟踪不同细胞群体随时间的出现，并通过原位蛋白表达验证我们的转录组数据。这些观察结果表明，移植的 HIO 确实再现了早期肠道发育，这巩固了它们作为人类肠道模型系统的价值。
