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首页> 外文期刊>Materials science & engineering >Thermoresponsive poly(N-isopropylacrylamide) hydrogel substrates micropatterned with poly(ethylene glycol) hydrogel for adipose mesenchymal stem cell spheroid formation and retrieval
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Thermoresponsive poly(N-isopropylacrylamide) hydrogel substrates micropatterned with poly(ethylene glycol) hydrogel for adipose mesenchymal stem cell spheroid formation and retrieval

机译：热响应聚（N-异丙基丙烯酰胺）水凝胶基材用聚（乙二醇）水凝胶用于脂肪间充质干细胞球体形成和检索

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Cell spheroid formation is necessary to develop three-dimensional (3D) cellular environments that provide appropriate cell-cell and cell-matrix interactions similar to in vivo environments without additional substrates. Although some methods including stirring culture, low adhesion plate culture, hanging drop, and microfluidics are used to construct cell spheroids, there is no method to fulfill all of the mass production of uniform spheroids, simple media change, and easy retrievability. Here, bulk poly(N-isopropylacrylamide) (PNIPAAm) hydrogel substrate (PHS) was used to fabricate, culture, and retrieve cell spheroids. Adipose-derived stem cells (ASCs) were cultured on bulk PHS to form spheroids. ASCs formed cell spheroids directly on substrates without additional manipulation. These spheroids adhered to the semi-adhesive substrate, while the spheroids fabricated using the nonadhesive surface method floated without getting fixed to the surface. Bulk PHS stiffness was evaluated using the compressive test (compressive modulus: 153 +/- 11 kPa). A poly(ethylene glycol) (PEG) hydrogel microwell pattern was created on PHS to control the spheroid size, forming uniform ASC spheroids between 100 and 150 mu m in diameter on 200 and 300 mu m well-patterned substrates. Cell-cell interactions in the resulting ASC spheroids were evaluated based on fibronectin and laminin expression; fluorescence intensifies of fibronectin- and laminin-immunostained images of ASC spheroids were 10.9 and 7.3 times higher than those of ASCs cultured on the tissue culture plate, respectively. ASC spheroids were detached following incubation at 4 degrees C for 10 min (retrieval efficiency: 74 +/- 19%). Retrieved spheroid cell viability was over 97.5%. The PEG hydrogel microwell-patterned PHS is a convenient spheroid fabrication and retrieval platform that can increase cell spheroid usage.
机译：细胞球体形成是开发三维（3D）细胞环境，其提供适当的细胞单元和细胞 - 矩阵相互作用，其在没有额外的基板的情况下类似于体内环境。虽然包括搅拌培养，低粘附板培养，悬挂液滴和微流体的一些方法用于构建细胞球体，但没有方法可以实现均匀球体的所有批量生产，简单的介质变化，易于检索性。这里，使用批量聚（N-异丙基丙烯酰胺）（PNIPAAM）水凝胶底物（pHS）来制造，培养和检索细胞球体。脂肪衍生的干细胞（ASCS）培养在本体pH上以形成球状体。 ASC在没有额外操纵的基板上直接形成细胞球体。这些球形粘附到半粘合剂基板上，同时使用不固定到表面的非粘性表面方法制造的球状体。使用压缩试验（压缩模量：153 +/- 11kPa）评估批量pHS僵硬。在pHS上产生聚（乙二醇）（PEG）水凝胶微孔图案以控制球状体尺寸，在200和300μm的直径上形成均匀的ASC球体在200和300μm的粗底板上。基于纤连蛋白和层状蛋白表达评估所得ASC球状体中的细胞细胞相互作用;纤维连接蛋白和层粘连蛋白 - 免疫染色图像的荧光分别比在组织培养板上培养的ASCS高10.9和7.3倍。在4摄氏度下孵育10分钟（检索效率：74 +/- 19％）后分离。检索到的球形细胞活力超过97.5％。 PEG水凝胶微孔图案化pHS是一种方便的球形制造和检索平台，可以增加细胞球体使用。

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