de Souza Isisdoris Rodrigues, Suzukawa Andreia Akemi, da Silva Horinouchi Cintia Delai, de Aguiar Alessandra Melo, Dallagiovanna Bruno
Laboratório de Biologia Básica de Células-Tronco, FIOCRUZ, Rua Professor Algacyr Munhoz Mader, 3775, Instituto Carlos Chagas, Curitiba, Paraná PR 81350-010 Brazil.
Present Address: Laboratório Nacional de Biociências Do Centro Nacional de Pesquisa Em Energia e Materiais (LNBIO-CNPEM) - Grupo de Engenharia Tecidual, Rua Giuseppe Máximo Scolfaro, 10000 - Polo II de Alta Tecnologia, Campinas, SP 13083-970 Brazil.
In Vitro Model. 2024 Oct 9;3(4-6):169-182. doi: 10.1007/s44164-024-00076-1. eCollection 2024 Dec.
Obesity is associated with several comorbidities that cause high mortality rates worldwide. Thus, the study of adipose tissue (AT) has become a target of high interest because of its crucial contribution to many metabolic diseases and metabolizing potential. However, many AT-related physiological, pathophysiological, and toxicological mechanisms in humans are still poorly understood, mainly due to the use of non-human animal models. Organ-on-chip (OoC) platform is a promising alternative to animal models. However, the use of adipose-derived human mesenchymal stem cells (hASCs) in these models is still scarce, and more knowledge on the effects properties of culturing hASCs in OoC models is needed. Here, we present the development of an OoC using hASCs to assess adipogenic differentiation. The device capability to increase hASC differentiation levels was confirmed by Nile red staining to verify lipid droplets inside cells after 10 days of culture and fluid flow of 10 µL/h. The Adipo-on-a-chip system increases hASC proliferation and differentiation area compared with the standard culture method under static conditions (96-well plates) verified in hASCs from different donors by image analysis of cells stained with Nile red. The expression of the gene is lower in the MPS, which calls attention to different homeostasis and control of lipids in cells in the MPS, compared with the plates. An increase of hASC proliferation in the MPS related to the 96-well plate was verified through protein Ki-67 expression. Cell and nuclei morphology (area, roundness, perimeter, width, length, width to length rate, symmetry, compactness, axial and radial properties to nuclei, and texture) and dominant direction of cells inside the MPS were evaluated to characterize hASCs in the present model. The presented microphysiological system (MPS) provides a promising tool for applications in mechanistic research aiming to investigate adipogenesis in AT and toxicological assessment based on the hASC differentiation potential.
肥胖与多种合并症相关，这些合并症在全球范围内导致了高死亡率。因此，由于脂肪组织（AT）对许多代谢性疾病的关键作用及其代谢潜力，对其研究已成为备受关注的目标。然而，人类中许多与AT相关的生理、病理生理和毒理学机制仍知之甚少，这主要是由于使用了非人类动物模型。芯片器官（OoC）平台是动物模型的一个有前景的替代方案。然而，在这些模型中使用脂肪来源的人间充质干细胞（hASC）仍然很少见，并且需要更多关于在OoC模型中培养hASC的效果和特性的知识。在这里，我们展示了一种使用hASC来评估成脂分化的OoC的开发。通过尼罗红染色证实了该装置在培养10天且流体流速为10 μL/h后增加hASC分化水平的能力，以验证细胞内的脂滴。与在静态条件下（96孔板）的标准培养方法相比，通过对用尼罗红染色的细胞进行图像分析，芯片上脂肪系统增加了来自不同供体的hASC的增殖和分化面积。与平板相比，MPS中该基因的表达较低，这引起了对MPS中细胞内脂质不同稳态和控制的关注。通过蛋白质Ki-67的表达证实了与96孔板相比，MPS中hASC增殖的增加。评估了MPS内细胞和细胞核的形态（面积、圆度、周长、宽度、长度、宽长比、对称性、紧凑性、细胞核的轴向和径向特性以及纹理）以及细胞的主导方向，以表征本模型中的hASC。所展示的微生理系统（MPS）为旨在研究AT中的脂肪生成和基于hASC分化潜力的毒理学评估的机制研究应用提供了一个有前景的工具。
