Human Adipose Derived Cells in Two- and Three-Dimensional Cultures: Functional Validation of an In Vitro Fat Construct
Human Adipose Derived Cells in Two- and Three-Dimensional Cultures: Functional Validation of an In Vitro Fat Construct
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DOI：
10.1155/2020/4242130
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发表时间：
2020-06-10
影响因子：
4.3
通讯作者：
Frazier, Trivia
中科院分区：
文献类型：
--
作者：
Bender, Robert;McCarthy, Michelle;Frazier, Trivia
肥胖，定义为身体质量指数30 kg/m2或以上，在美国和全球范围内的发病率和频率显著增加。相关的合并症包括心血管疾病、2型糖尿病、代谢综合征和非酒精性脂肪肝疾病，这使得人们关注促进肥胖预防和治疗的机制。通常使用的体外模型采用人或小鼠前脂肪细胞系的二维（2D）格式。由于这些模型的结构、生物化学和生物学限制，越来越多的注意力被放在用于三维（3D）培养的“芯片上器官”技术上。在此，我们描述了一种采用冷冻保存的原代人基质血管部分（SVF）细胞和人血液制品衍生的生物支架在体外产生3D脂肪库的方法。基于增殖、流式细胞术、脂肪形成分化、共聚焦显微镜/免疫荧光和功能测定（脂肪因子分泌、葡萄糖摄取和脂解），相对于2D培养物，“芯片上脂肪”3D培养物已经得到验证。因此，体外培养系统证明了人源化3D白色脂肪组织（WAT）模型所需的关键特征。
Obesity, defined as a body mass index of 30 kg/m(2)or above, has increased considerably in incidence and frequency within the United States and globally. Associated comorbidities including cardiovascular disease, type 2 diabetes mellitus, metabolic syndrome, and nonalcoholic fatty liver disease have led to a focus on the mechanisms promoting the prevention and treatment of obesity. Commonly utilizedin vitromodels employ human or mouse preadipocyte cell lines in a 2-dimensional (2D) format. Due to the structural, biochemical, and biological limitations of these models, increased attention has been placed on "organ on a chip" technologies for a 3-dimensional (3D) culture. Herein, we describe a method employing cryopreserved primary human stromal vascular fraction (SVF) cells and a human blood product-derived biological scaffold to create a 3D adipose depot in vitro. The "fat-on-chip" 3D cultures have been validated relative to 2D cultures based on proliferation, flow cytometry, adipogenic differentiation, confocal microscopy/immunofluorescence, and functional assays (adipokine secretion, glucose uptake, and lipolysis). Thus, thein vitroculture system demonstrates the critical characteristics required for a humanized 3D white adipose tissue (WAT) model.