Forget A, Burzava A L S, Delalat B, Vasilev K, Harding F J, Blencowe A, Voelcker N H
Science and Engineering Faculty, Queensland University of Technology, Brisbane, QLD 4001, Australia and Cooperative Research Centre for Cell Therapy Manufacturing (CRC-CTM), Adelaide 5000, Australia.
Cooperative Research Centre for Cell Therapy Manufacturing (CRC-CTM), Adelaide 5000, Australia and Monash Institute of Pharmaceutical Sciences, Monash University, Parkville, VIC 3052, Australia.
Biomater Sci. 2017 Mar 28;5(4):828-836. doi: 10.1039/c6bm00916f.
Cell aggregates reproduce many features of the natural architecture of functional tissues, and have therefore become an important in vitro model of tissue function. In this study, we present an efficient and rapid method for the fabrication of site specific functionalised poly(dimethylsiloxane) (PDMS) microwell arrays that promote the formation of insulin-producing beta cell (MIN6) aggregates. Microwells were prepared using an ice templating technique whereby aqueous droplets were frozen on a surface and PDMS was cast on top to form a replica. By employing an aqueous alkali hydroxide solution, we demonstrate exclusive etching and functionalisation of the microwell inner surface, thereby allowing the selective absorption of biological factors within the microwells. Additionally, by manipulating surface wettability of the substrate through plasma polymer coating, the shape and profile of the microwells could be tailored. Microwells coated with antifouling Pluronic 123, bovine serum albumin, collagen type IV or insulin growth factor 2 were employed to investigate the formation and stability of MIN6 aggregates in microwells of different shapes. MIN6 aggregates formed with this technique retained insulin expression. These results demonstrate the potential of this platform for the rapid screening of biological factors influencing the formation and response of insulin-producing cell aggregates without the need for expensive micromachining techniques.
细胞聚集体重现了功能组织自然结构的许多特征，因此已成为组织功能的重要体外模型。在本研究中，我们提出了一种高效快速的方法来制造位点特异性功能化的聚二甲基硅氧烷（PDMS）微孔阵列，该阵列可促进产生胰岛素的β细胞（MIN6）聚集体的形成。微孔是使用冰模板技术制备的，即将水滴冷冻在表面上，然后在顶部浇铸PDMS以形成复制品。通过使用氢氧化碱水溶液，我们展示了微孔内表面的选择性蚀刻和功能化，从而允许在微孔内选择性吸收生物因子。此外，通过等离子体聚合物涂层控制基材的表面润湿性，可以调整微孔的形状和轮廓。使用涂有抗污剂普朗尼克123、牛血清白蛋白、IV型胶原蛋白或胰岛素生长因子2的微孔来研究不同形状微孔中MIN6聚集体的形成和稳定性。用该技术形成的MIN6聚集体保留了胰岛素表达。这些结果证明了该平台在快速筛选影响产生胰岛素细胞聚集体形成和反应的生物因子方面的潜力，而无需昂贵的微加工技术。
