In vitro generation of human pluripotent stem cell derived lung organoids.
In vitro generation of human pluripotent stem cell derived lung organoids.
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DOI：
10.7554/elife.05098
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发表时间：
2015-03-24
期刊：
影响因子：
7.7
通讯作者：
Spence JR
中科院分区：
文献类型：
--
作者：
Dye BR;Hill DR;Ferguson MA;Tsai YH;Nagy MS;Dyal R;Wells JM;Mayhew CN;Nattiv R;Klein OD;White ES;Deutsch GH;Spence JR
最近在许多器官系统的三维（3D）类器官培养方面的突破已经导致了新的生理复杂的体外模型来研究人类发育和疾病。在这里，我们报告了人类多能干细胞（hPSC）（胚胎和诱导）逐步分化为肺类器官。通过操纵发育信号传导途径，hPSC产生腹前肠球状体，然后将其扩增成人肺类器官（HLO）。HLO由肺的上皮和间充质隔室组成，其组织结构特征与天然肺相似。HLO具有上气道样上皮，其具有被平滑肌和肌成纤维细胞包围的基底细胞和未成熟纤毛细胞，以及具有适当细胞类型的肺泡样结构域。使用RNA测序，我们表明，HLO是非常相似的人胎肺的基础上的全球转录谱，这表明HLO是一个很好的模型来研究人类肺的发育，成熟和疾病。DOI：http://dx.doi.org/10.7554/eLife.05098.001细胞行为传统上是在实验室的二维环境中研究的，细胞在细胞培养皿中以薄层生长。然而，人体中的大多数细胞作为复杂组织和器官的一部分存在于三维环境中，因此研究人员一直试图在实验室中重建这些环境。到目前为止，已经成功地生成了几个这样的“类器官”，包括人类肠道，胃，大脑和肝脏的模型。这些类器官可以模拟真实的组织的反应，并可用于研究器官如何形成、随着疾病而变化以及它们如何对潜在疗法做出反应。在这里，Dye等人通过诱导人类干细胞成为特定类型的细胞，然后在培养皿中形成复杂的组织，开发了一种新的人类肺部三维模型。为了制造这些肺类器官，Dye等人操纵了几种在动物胚胎发育过程中控制器官形成的信号通路。首先，干细胞被指示形成一种称为内胚层的组织，这种组织存在于早期胚胎中，并产生肺，肝脏和其他几个内部器官。然后，Dye等人激活了两条重要的发育途径，已知这两条途径使内胚层形成三维肠组织。然而，通过同时抑制其他两个关键的发育途径，内胚层变成了类似于胚胎中发现的早期肺的组织。这种早期的肺样组织在发育过程中形成了三维的球形结构。下一个挑战是使这些结构发育成肺组织。Dye等人研究出了一种方法来做到这一点，该方法包括将细胞暴露于参与肺发育的其他蛋白质。由此产生的肺类器官在实验室培养中存活了100多天，并发育成组织良好的结构，其中包含许多在肺中发现的细胞类型。进一步的分析显示，肺类器官中的基因活性类似于发育中的人类胎儿的肺，这表明在培养皿中生长的肺类器官尚未完全成熟。Dye等人的发现为在培养中创造人类肺部类器官提供了一种新方法，可能为研究肺部发育和疾病开辟新途径。DOI：http://dx.doi.org/10.7554/eLife.05098.002网站
Recent breakthroughs in 3-dimensional (3D) organoid cultures for many organ systems have led to new physiologically complex in vitro models to study human development and disease. Here, we report the step-wise differentiation of human pluripotent stem cells (hPSCs) (embryonic and induced) into lung organoids. By manipulating developmental signaling pathways hPSCs generate ventral-anterior foregut spheroids, which are then expanded into human lung organoids (HLOs). HLOs consist of epithelial and mesenchymal compartments of the lung, organized with structural features similar to the native lung. HLOs possess upper airway-like epithelium with basal cells and immature ciliated cells surrounded by smooth muscle and myofibroblasts as well as an alveolar-like domain with appropriate cell types. Using RNA-sequencing, we show that HLOs are remarkably similar to human fetal lung based on global transcriptional profiles, suggesting that HLOs are an excellent model to study human lung development, maturation and disease. DOI: http://dx.doi.org/10.7554/eLife.05098.001 Cell behavior has traditionally been studied in the lab in two-dimensional situations, where cells are grown in thin layers on cell-culture dishes. However, most cells in the body exist in a three-dimensional environment as part of complex tissues and organs, and so researchers have been attempting to re-create these environments in the lab. To date, several such ‘organoids’ have been successfully generated, including models of the human intestine, stomach, brain and liver. These organoids can mimic the responses of real tissues and can be used to investigate how organs form, change with disease, and how they might respond to potential therapies. Here, Dye et al. developed a new three-dimensional model of the human lung by coaxing human stem cells to become specific types of cells that then formed complex tissues in a petri dish. To make these lung organoids, Dye et al. manipulated several of the signaling pathways that control the formation of organs during the development of animal embryos. First, the stem cells were instructed to form a type of tissue called endoderm, which is found in early embryos and gives rise to the lung, liver and other several other internal organs. Then, Dye et al. activated two important developmental pathways that are known to make endoderm form three-dimensional intestinal tissue. However, by inhibiting two other key developmental pathways at the same time, the endoderm became tissue that resembles the early lung found in embryos instead. This early lung-like tissue formed three-dimensional spherical structures as it developed. The next challenge was to make these structures develop into lung tissue. Dye et al. worked out a method to do this, which involved exposing the cells to additional proteins that are involved in lung development. The resulting lung organoids survived in laboratory cultures for over 100 days and developed into well-organized structures that contain many of the types of cells found in the lung. Further analysis revealed the gene activity in the lung organoids resembles that of the lung of a developing human fetus, suggesting that lung organoids grown in the dish are not fully mature. Dye et al.'s findings provide a new approach for creating human lung organoids in culture that may open up new avenues for investigating lung development and diseases. DOI: http://dx.doi.org/10.7554/eLife.05098.002