Enhanced Adipogenic Differentiation of Human Adipose-Derived Stem Cells in an In Vitro Microenvironment: The Preparation of Adipose-Like Microtissues Using a Three-Dimensional Culture
Enhanced Adipogenic Differentiation of Human Adipose-Derived Stem Cells in an In Vitro Microenvironment: The Preparation of Adipose-Like Microtissues Using a Three-Dimensional Culture
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DOI：
10.3727/215517916x693096
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发表时间：
2016-01-01
期刊：
影响因子：
--
通讯作者：
Hayashi, Shuji
中科院分区：
文献类型：
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作者：
Miyamoto, Yoshitaka;Ikeuchi, Masashi;Hayashi, Shuji
近年来，干细胞在细胞治疗中的应用得到了广泛的研究。在各种类型的干细胞中，人脂肪组织源性干细胞（human adipose tissue-derived stem cells, ASCs）可以以相对较少的传代量获得，并且具有稳定的质量。ASCs可以分化为多种细胞类型，如脂肪细胞和外胚层细胞。因此，我们专注于这种分化所需的体外微环境，并试图利用微机电系统诱导人类干细胞向微组织分化。我们首先在三维（3D）培养中评估了人类ASC球体的成脂分化。然后，我们使用3D组合TASCL装置创建体外微环境，并试图诱导人ASCs的成脂分化。在维持培养基和脂肪细胞分化培养基中培养的人ASC球体，通过脂滴油红O染色，根据积累的甘油三酯的数量来评估其分化程度。在两种培养基中均证实了分化，但3D培养的人ASCs中甘油三酯含量高于2D培养。在较短的培养时间内，3D培养比2D培养观察到更大的成脂分化。与2D培养或使用维持培养基的培养相比，使用TASCL装置与成脂分化培养基的3D培养促进了人类ASCs向成脂谱系的更大分化。总之，TASCL装置创造了一个好客的体外微环境，因此可能是在3D培养中诱导分化的有用工具。由此产生的人类ASC球体是“脂肪样微组织”，形成完美的球形聚集，有望应用于再生医学和细胞移植。
The application of stem cells for cell therapy has been extensively studied in recent years. Among the various types of stem cells, human adipose tissue-derived stem cells (ASCs) can be obtained in large quantities with relatively few passages, and they possess a stable quality. ASCs can differentiate into a number of cell types, such as adipose cells and ectodermal cells. We therefore focused on the in vitro microenvironment required for such differentiation and attempted to induce the differentiation of human stem cells into microtissues using a microelectromechanical system. We first evaluated the adipogenic differentiation of human ASC spheroids in a three-dimensional (3D) culture. We then created the in vitro microenvironment using a 3D combinatorial TASCL device and attempted to induce the adipogenic differentiation of human ASCs. The differentiation of human ASC spheroids cultured in maintenance medium and those cultured in adipocyte differentiation medium was evaluated via Oil red O staining using lipid droplets based on the quantity of accumulated triglycerides. The differentiation was confirmed in both media, but the human ASCs in the 3D cultures contained higher amounts of triglycerides than those in the 2D cultures. In the short culture period, greater adipogenic differentiation was observed in the 3D cultures than in the 2D cultures. The 3D culture using the TASCL device with adipogenic differentiation medium promoted greater differentiation of human ASCs into adipogenic lineages than either a 2D culture or a culture using a maintenance medium. In summary, the TASCL device created a hospitable in vitro microenvironment and may therefore be a useful tool for the induction of differentiation in 3D culture. The resultant human ASC spheroids were "adipose-like microtissues" that formed spherical aggregation perfectly and are expected to be applicable in regenerative medicine as well as cell transplantation.