Torisawa Yu-suke, Takagi Airi, Nashimoto Yuji, Yasukawa Tomoyuki, Shiku Hitoshi, Matsue Tomokazu
Graduate School of Environmental Studies, Tohoku University, Sendai 980-8579, Japan.
Biomaterials. 2007 Jan;28(3):559-66. doi: 10.1016/j.biomaterials.2006.08.054. Epub 2006 Sep 20.
We describe a novel multicellular spheroid culture system that facilitates the easy preparation and culture of a spheroid microarray for the long-term monitoring of cellular activity. A spheroid culture device with an array of pyramid-like microholes was constructed in a silicon chip that was equipped with elastomeric microchannels. A cell suspension was introduced via the microfluidic channel into the microstructure that comprised silicon microholes and elastomeric microwells. A single spheroid can be formed and localized precisely within each microstructure. Since the culture medium could be replaced via the microchannels, a long-term culture (of approximately 2 weeks) is available on the chip. Measurement of albumin production in the hepatoma cell line (HepG2) showed that the liver-specific functions were maintained for 2 weeks. Based on the cellular respiratory activity, the cellular viability of the spheroid array on the chip was evaluated using scanning electrochemical microscopy. Responses to four different chemical stimulations were simultaneously detected on the same chip, thus demonstrating that each channel could be evaluated independently under various stimulation conditions. Our spheroid culture system facilitated the understanding of spheroid formation, culture, and viability assay on a single chip, thus functioning as a useful drug-screening device for cancer and liver cells.
我们描述了一种新型的多细胞球体培养系统，该系统便于轻松制备和培养球体微阵列，用于长期监测细胞活性。在配备有弹性微通道的硅芯片中构建了一种具有金字塔状微孔阵列的球体培养装置。通过微流体通道将细胞悬液引入由硅微孔和弹性微阱组成的微结构中。单个球体可以在每个微结构中精确形成并定位。由于可以通过微通道更换培养基，因此可以在芯片上进行长期培养（约2周）。对肝癌细胞系（HepG2）中白蛋白产生的测量表明，肝脏特异性功能维持了2周。基于细胞呼吸活性，使用扫描电化学显微镜评估芯片上球体阵列的细胞活力。在同一芯片上同时检测对四种不同化学刺激的反应，从而证明每个通道可以在各种刺激条件下独立评估。我们的球体培养系统有助于在单个芯片上理解球体的形成、培养和活力测定，从而作为一种用于癌症和肝细胞的有用药物筛选装置。
