开发和分析无支架脂肪球。

IF 3.5 4区 生物学 Q2 ENDOCRINOLOGY & METABOLISM
Adipocyte Pub Date : 2024-12-01 Epub Date: 2024-06-12 DOI:10.1080/21623945.2024.2347215
Jesse Liszewski, Aloysious Klingelhutz, Edward A Sander, James Ankrum
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引用次数: 0

摘要

脂肪组织在代谢综合征、自身免疫性疾病和许多癌症中起着至关重要的作用。由于脂肪在人类健康的许多方面都起着重要作用,因此亟需能复制脂肪结构和功能的体外模型。传统的单层模型虽然方便,但也有局限性,与三维模型相比显示出异质性和功能差异。单层培养的脂肪细胞难以脱落、分化效率低,而三维培养的健康脂肪细胞则能积聚大量脂滴、分泌脂肪连素并产生低水平的炎症细胞因子。从单层模型到更复杂的三维模型的转变旨在更好地复制健康脂肪组织的培养生理。本研究介绍了一种使用无支架球体模型生成脂肪器官组织的简单易行的方案。与以前的技术相比,该方法利用 96 孔球形板或琼脂糖微模,提高了产量、均匀性和易用性。该方案可用于多种应用,包括药物测试、毒素筛选、组织工程学和共培养。这两种方法的选择取决于实验目标,96 孔板提供了个性化控制,而微模具则提供了规模优势。概述的方案包括基质血管部分细胞的分离、扩增和表征,随后是球形体形成的详细步骤和可选的下游分析。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Development and analysis of scaffold-free adipose spheroids.

Adipose tissue plays a crucial role in metabolic syndrome, autoimmune diseases, and many cancers. Because of adipose's role in so many aspects of human health, there is a critical need for in vitro models that replicate adipose architecture and function. Traditional monolayer models, despite their convenience, are limited, showing heterogeneity and functional differences compared to 3D models. While monolayer cultures struggle with detachment and inefficient differentiation, healthy adipocytes in 3D culture accumulate large lipid droplets, secrete adiponectin, and produce low levels of inflammatory cytokines. The shift from monolayer models to more complex 3D models aims to better replicate the physiology of healthy adipose tissue in culture. This study introduces a simple and accessible protocol for generating adipose organoids using a scaffold-free spheroid model. The method, utilizing either 96-well spheroid plates or agarose micromolds, demonstrates increased throughput, uniformity, and ease of handling compared to previous techniques. This protocol allows for diverse applications, including drug testing, toxin screening, tissue engineering, and co-culturing. The choice between the two methods depends on the experimental goals, with the 96-well plate providing individualized control and the micromold offering scale advantages. The outlined protocol covers isolation, expansion, and characterization of stromal vascular fraction cells, followed by detailed steps for spheroid formation and optional downstream analyses.

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来源期刊
Adipocyte
Adipocyte Medicine-Histology
CiteScore
6.50
自引率
3.00%
发文量
46
审稿时长
32 weeks
期刊介绍: Adipocyte recognizes that the adipose tissue is the largest endocrine organ in the body, and explores the link between dysfunctional adipose tissue and the growing number of chronic diseases including diabetes, hypertension, cardiovascular disease and cancer. Historically, the primary function of the adipose tissue was limited to energy storage and thermoregulation. However, a plethora of research over the past 3 decades has recognized the dynamic role of the adipose tissue and its contribution to a variety of physiological processes including reproduction, angiogenesis, apoptosis, inflammation, blood pressure, coagulation, fibrinolysis, immunity and general metabolic homeostasis. The field of Adipose Tissue research has grown tremendously, and Adipocyte is the first international peer-reviewed journal of its kind providing a multi-disciplinary forum for research focusing exclusively on all aspects of adipose tissue physiology and pathophysiology. Adipocyte accepts high-profile submissions in basic, translational and clinical research.
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