Vascularized adipocyte organoid model using isolated human microvessel fragments
Vascularized adipocyte organoid model using isolated human microvessel fragments
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DOI：
10.1088/1758-5090/abe187
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发表时间：
2021-07-01
期刊：
影响因子：
9
通讯作者：
Hoying, James B.
中科院分区：
文献类型：
--
作者：
Strobel, Hannah A.;Gerton, Thomas;Hoying, James B.
组织类器官被证明对于在各种应用中建模组织健康和疾病是有价值的。这部分是由于在3D组织样空间内培养的动态细胞-细胞相互作用。为此，类器官越多地再现天然组织中发现的不同细胞-细胞相互作用，例如实质和微血管之间的相互作用，模型的保真度就越好。微脉管系统由一系列细胞类型组成，不仅提供灌注以支持组织健康，而且提供重要的细胞相互作用和生物化学动力学，这对组织表型和功能很重要。在这里，我们将从脂肪组织分离的完整的人微血管片段整合到类器官中，以形成间充质干细胞（MSC）和脂肪细胞血管化的类器官。分离的微血管保留其天然结构和细胞组成，提供了类器官内微血管系统的更完整表示。当置于3D胶原基质中时，通过在由MSC或MSC衍生的脂肪细胞组成的类器官内的萌芽血管生成而扩张的微血管从类器官中生长出来。在MSC类器官中，50个MSC与1个微血管片段的比例产生了最佳的血管化反应。我们开发了一种新的分化方案，使间充质干细胞分化成脂肪细胞，同时促进微血管生成。脂肪细胞类器官含有血管网络，在脂解试验中有反应，并表达功能性脂肪细胞标志物脂联素和过氧化物酶体增殖物激活受体γ。微血管的存在促进了脂肪细胞胰岛素受体的表达和肿瘤坏死因子α刺激后白细胞介素-6的分泌。总的来说，我们证明了一种强大的方法，用于血管化高细胞密度的类器官，对其他组织也有潜在的影响。
Tissue organoids are proving valuable for modeling tissue health and disease in a variety of applications. This is due, in part, to the dynamic cell-cell interactions fostered within the 3D tissue-like space. To this end, the more that organoids recapitulate the different cell-cell interactions found in native tissue, such as that between parenchyma and the microvasculature, the better the fidelity of the model. The microvasculature, which is comprised of a spectrum of cell types, provides not only perfusion in its support of tissue health, but also important cellular interactions and biochemical dynamics important in tissue phenotype and function. Here, we incorporate whole, intact human microvessel fragments isolated from adipose tissue into organoids to form both mesenchymal stem cell (MSC) and adipocyte vascularized organoids. Isolated microvessels retain their native structure and cell composition, providing a more complete representation of the microvasculature within the organoids. Microvessels expanded via sprouting angiogenesis within organoids comprised of either MSCs or MSC-derived adipocytes grew out of the organoids when placed in a 3D collagen matrix. In MSC organoids, a ratio of 50 MSCs to 1 microvessel fragment created the optimal vascularization response. We developed a new differentiation protocol that enabled the differentiation of MSCs into adipocytes while simultaneously promoting microvessel angiogenesis. The adipocyte organoids contained vascular networks, were responsive in a lipolysis assay, and expressed the functional adipocyte markers adiponectin and peroxisome proliferator-activated receptor gamma. The presence of microvessels promoted insulin receptor expression by adipocytes and modified interleukin-6 secretion following a tumor necrosis factor alpha challenge. Overall, we demonstrate a robust method for vascularizing high cell-density organoids with potential implications for other tissues as well.