PDF(Pubmed)
Abstract：
Adipose tissue is a complex and multifaceted endocrine organ located throughout the body. The dysfunction of adipose tissue is known to induce a wide variety of comorbidities that can negatively impact one\'s health and quality of life. In addition to behavioral changes, drugs that target dysfunctional adipose tissue to treat associated diseases are clinically needed. Regarding drug-testing platforms, animal models are the most popular models, limited by known differences from humans in genetics and physiology. Two-dimensional and static three-dimensional (3D) cell cultures are also used. Still, these in vitro models with static culture fail to recapitulate the phenotype and function of adipocytes seen in vivo. To combat this, our lab has developed an adipose tissue microphysiological system. A perfusion bioreactor with dual-flow chambers is 3D printed, which enables individualized top and bottom medium flows after adipose tissues are inserted as a barrier. Human progenitor cells, such as human mesenchymal stem cells, are embedded within a gelatin scaffold and in situ adipogenic differentiation within the bioreactor. Medium flow is established via a syringe pump system, allowing in vivo-like conditions to be maintained. The novel bioreactor-cultured adipose tissues represent a versatile disease modeling and drug-testing system. Here, we present the step-by-step methods to generate the bioreactors and adipose tissues. We also show the process of collecting and analyzing samples. In addition, we highlight the critical steps that require particular attention in notes.
摘要：
脂肪组织是位于全身的复杂和多方面的内分泌器官。已知脂肪组织的功能障碍会引起多种合并症，这些合并症会对人的健康和生活质量产生负面影响。除了行为上的改变,临床上需要针对功能失调的脂肪组织的药物来治疗相关疾病。关于药物测试平台，动物模型是最受欢迎的模型，受限于与人类在遗传学和生理学方面的已知差异。还使用二维和静态三维(3D)细胞培养物。尽管如此，这些静态培养的体外模型未能概括体内脂肪细胞的表型和功能。为了解决这个问题，我们的实验室开发了一种脂肪组织微生理系统。带有双流室的灌注生物反应器是3D打印的，这使得在插入脂肪组织作为屏障后能够实现个性化的顶部和底部培养基流动。人类祖细胞，例如人类间充质干细胞，包埋在明胶支架内并在生物反应器内原位成脂分化。介质流通过注射泵系统建立，允许维持体内样条件。新型生物反应器培养的脂肪组织代表了一种通用的疾病建模和药物测试系统。这里,我们介绍了逐步生成生物反应器和脂肪组织的方法。我们还展示了收集和分析样品的过程。此外,我们强调了需要特别注意的关键步骤。
