Highly-parallel production of designer organoids by mosaic patterning of progenitors.
Highly-parallel production of designer organoids by mosaic patterning of progenitors.
复制标题
通过祖细胞的马赛克图案高度并行生产设计师类器官。
DOI：
10.1101/2023.10.25.564017
复制
发表时间：
2023
期刊：
影响因子：
--
通讯作者：
Hughes,AlexJ
中科院分区：
文献类型：
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作者：
Porter,CatherineM;Qian,GraceC;Grindel,SamuelH;Hughes,AlexJ
源自人类干细胞的类器官是疾病建模、再生医学和基础研究的一种有前景的方法。然而，类器官的变异性和对形态结果的有限控制仍然是挑战。一个悬而未决的问题是，对培养条件的工程控制可以在多大程度上引导类器官形成特定的成分。在这里，我们扩展了 DNA“尼龙搭扣”细胞图案化方法，精确控制微孔阵列内促成肾单位祖细胞 (NP) 类器官和嵌合 NP/输尿管芽 (UB) 尖端细胞类器官的人类诱导多能干细胞来源的祖细胞的数量和比例。我们展示了对类器官大小和形态的长期控制，不受几何约束的影响。然后，我们展示了依赖于初始祖细胞组成的类器官组织比例的新趋势。这些包括与仅 NP 类器官相比，镶嵌 NP/UB 类器官中肾单位和基质细胞的代表性更高，以及镶嵌类器官中“金发姑娘”初始细胞比例，可优化近端小管结构的形成。
Organoids derived from human stem cells are a promising approach for disease modeling, regenerative medicine, and fundamental research. However, organoid variability and limited control over morphological outcomes remain as challenges. One open question is the extent to which engineering control over culture conditions can guide organoids to specific compositions. Here, we extend a DNA "velcro" cell patterning approach, precisely controlling the number and ratio of human induced pluripotent stem cell-derived progenitors contributing to nephron progenitor (NP) organoids and mosaic NP/ureteric bud (UB) tip cell organoids within arrays of microwells. We demonstrate long-term control over organoid size and morphology, decoupled from geometric constraints. We then show emergent trends in organoid tissue proportions that depend on initial progenitor cell composition. These include higher nephron and stromal cell representation in mosaic NP/UB organoids vs. NP-only organoids and a "goldilocks" initial cell ratio in mosaic organoids that optimizes the formation of proximal tubule structures.