Department of Life and Environment Engineering, The University of Kitakyushu, 1-1 Hibikino, Wakamatsu-ku, Kitakyushu 808-0135, Japan.
J Biosci Bioeng. 2010 Aug;110(2):223-9. doi: 10.1016/j.jbiosc.2010.01.021. Epub 2010 Feb 25.
The formation of three-dimensional cell microspheres such as spheroids, embryoid bodies, and neurospheres has attracted attention as a useful culture technique. In this study, we investigated a technique for effective cell microsphere production by using specially prepared microchip. The basic chip design was a multimicrowell structure in triangular arrangement within a 100-mm(2) region in the center of a polymethylmethacrylate (PMMA) plate (24x24 mm(2)), the surface of which was modified with polyethylene glycol (PEG) to render it nonadhesive to cells. We also designed six similar chips with microwell diameters of 200, 300, 400, 600, 800, and 1000 microm to investigate the effect of the microwell diameter on the cell microsphere diameter. Rat hepatocytes, HepG2 cells, mouse embryonic stem (ES) cells, and mouse neural progenitor/stem (NPS) cells formed hepatocyte spheroids, HepG2 spheroids, embryoid bodies, and neurospheres, respectively, in the microwells within 5 days of culture. For all the cells, a single microsphere was formed in each microwell under all the chip conditions, and such microsphere configurations remained throughout the culture period. Furthermore, the microsphere diameters of each type of cell were strongly positively correlated with the microwell diameters of the chips, suggesting that microsphere diameter can be factitiously controlled by using different chip conditions. Thus, this chip technique is a promising cellular platform for tissue engineering or regenerative medicine research, pharmacological and toxicological studies, and fundamental studies in cell biology.
三维细胞微球的形成，如球体、类胚体和神经球，作为一种有用的培养技术引起了关注。在这项研究中，我们研究了一种使用特殊制备的微芯片有效生产细胞微球的技术。基本芯片设计是在聚甲基丙烯酸甲酯（PMMA）板（24x24mm²）中心 100mm²区域内的三角形排列的多微井结构，其表面用聚乙二醇（PEG）修饰以使其对细胞无粘附性。我们还设计了六个类似的芯片，其微井直径分别为 200、300、400、600、800 和 1000μm，以研究微井直径对细胞微球直径的影响。大鼠肝细胞、HepG2 细胞、小鼠胚胎干细胞和小鼠神经祖细胞/干细胞分别在培养 5 天内在微井中形成肝细胞球体、HepG2 球体、类胚体和神经球。对于所有细胞，在所有芯片条件下，每个微井中均形成单个微球，并且在整个培养期间保持这种微球构型。此外，每种细胞的微球直径与芯片的微井直径呈强烈正相关，表明可以通过使用不同的芯片条件人为控制微球直径。因此，这种芯片技术是组织工程或再生医学研究、药理学和毒理学研究以及细胞生物学基础研究的有前途的细胞平台。
