Department of Surgery, University of Cincinnati, Cincinnati, OH; and; Division of Pediatric General and Thoracic Surgery, Cincinnati Children's Hospital Medical Center, Cincinnati, OH.
Division of Pediatric General and Thoracic Surgery, Cincinnati Children's Hospital Medical Center, Cincinnati, OH.
Surgery. 2018 Oct;164(4):643-650. doi: 10.1016/j.surg.2018.04.048. Epub 2018 Jul 30.
We previously described the development of human intestinal organoids from pluripotent stem cells, as well as their in vivo maturation when transplanted into the mouse kidney capsule. While sufficient for certain aspects of study, this model has limitations. Herein, we describe an alternative model of human intestinal organoids transplantation into the mouse mesentery. We hypothesize that efficient engraftment and marked differentiation of human intestinal organoids will be similar to our kidney model yet in a more anatomically appropriate location allowing for improved in vivo modeling.
Human intestinal organoids were generated by directed differentiation of H1 embryonic stem cells. Human intestinal organoids were then transplanted into the mesentery of immunosuppressed mice. Gross and histologic analysis of tissue was performed.
Human intestinal organoids were transplanted into the mouse mesentery and allowed to grow for 10 weeks. Mouse survival was 85%, and among the surviving mice, 82% of transplanted human intestinal organoids successfully engrafted. Upon graft harvest, transplanted HIOs were larger than in vitro human intestinal organoids (1.75 mm vs 6.27 mm, P < .0001) and grew along a vascular pedicle, allowing for interventions and reconstructive surgeries to access the human intestinal organoid lumen. Histologic analyses of transplanted human intestinal organoids confirmed the presence of major cell types, as well as stem cell activity.
The mouse mesentery is a viable location for the transplantation of human intestinal organoids, yielding grafts of reproducible size and quality. This improved model serves to advance functional and translational studies of human intestinal organoids.
我们之前描述了从多能干细胞中开发人类肠道类器官，以及将其移植到小鼠肾囊内后的体内成熟过程。虽然该模型足以满足某些研究方面的需求，但它仍存在一些局限性。在此，我们描述了一种将人类肠道类器官移植到小鼠肠系膜的替代模型。我们假设，人类肠道类器官的有效植入和显著分化将与我们的肾脏模型相似，但在更具解剖学意义的位置，从而可以更好地进行体内建模。
通过定向分化 H1 胚胎干细胞生成人类肠道类器官。然后将人类肠道类器官移植到免疫抑制小鼠的肠系膜中。对组织进行大体和组织学分析。
人类肠道类器官被移植到小鼠肠系膜中并允许生长 10 周。小鼠的存活率为 85%，在存活的小鼠中，82%的移植人类肠道类器官成功植入。在移植物收获时，移植的 HIO 比体外培养的人类肠道类器官更大（1.75 毫米比 6.27 毫米，P<.0001），并且沿着血管蒂生长，允许进行干预和重建手术以进入人类肠道类器官腔。移植的人类肠道类器官的组织学分析证实了主要细胞类型的存在以及干细胞活性。
小鼠肠系膜是移植人类肠道类器官的可行位置，可产生可重复大小和质量的移植物。这种改进的模型有助于推进人类肠道类器官的功能和转化研究。
