RIKEN Center for Biosystems Dynamics Research, Osaka, Japan.
Graduate School of Frontier Biosciences, Osaka University, Suita, Osaka, Japan.
PLoS One. 2020 May 20;15(5):e0232518. doi: 10.1371/journal.pone.0232518. eCollection 2020.
Patterned cell culturing is one of the most useful techniques for understanding the interaction between geometric conditions surrounding cells and their behaviors. The authors previously proposed a simple method for cell patterning with an agarose gel microstructure fabricated by microcasting with a degassed polydimethylsiloxane (PDMS) mold. Although the vacuum pressure produced from the degassed PDMS can drive a highly viscous agarose solution, the influence of solution viscosity on the casting process is unknown. This study investigated the influences of micro-channel dimensions or solution viscosity on the flow of the solution in a micro-channel of a PDMS mold by both experiments and numerical simulation. It was found experimentally that the degassed PDMS mold was able to drive a solution with a viscosity under 575 mPa·s. A simulation model was developed which can well estimate the flow rate in various dimensions of micro-channels. Cross-linked albumin has low viscosity (1 mPa·s) in aqueous solution and can undergo a one-way dehydration process from solution to solid that produces cellular repellency after dehydration. A microstructure of cross-linked albumin was fabricated on a cell culture dish by the microcasting method. After cells were seeded and cultivated on the cell culture dish with the microstructure for 7 days, the cellular pattern of mouse skeletal myoblast cell line C2C12 was observed. The microcasting with cross-linked albumin solution enables preparation of patterned cell culture systems more quickly in comparison with the previous agarose gel casting, which requires a gelation process before the dehydration process.
图案化细胞培养是理解细胞周围几何条件与其行为之间相互作用的最有用技术之一。作者先前提出了一种使用微铸造与脱气聚二甲基硅氧烷(PDMS)模具制造琼脂糖凝胶微结构的简单细胞图案化方法。尽管脱气 PDMS 产生的真空压力可以驱动高粘性的琼脂糖溶液,但溶液粘度对铸造过程的影响尚不清楚。本研究通过实验和数值模拟研究了微通道尺寸或溶液粘度对 PDMS 模具微通道中溶液流动的影响。实验发现,脱气 PDMS 模具能够驱动粘度低于 575 mPa·s 的溶液。开发了一个模拟模型,该模型可以很好地估计各种尺寸微通道中的流速。交联白蛋白在水溶液中的粘度低(1 mPa·s),并且可以从溶液到固体经历单向脱水过程,脱水后产生细胞排斥性。通过微铸造方法在细胞培养盘中制造交联白蛋白的微结构。在细胞培养盘上接种和培养细胞 7 天后,观察到小鼠成肌细胞系 C2C12 的细胞图案。与之前需要脱水过程前的凝胶化过程的琼脂糖凝胶铸造相比,交联白蛋白溶液的微铸造可以更快地制备图案化细胞培养系统。