Protein Aggregation Using Gold Nanoparticles and Organization between Patterned C2C12 Cells

自我組織行為(Self-organization)是一種在自然界中常見的現象。在生物領域中，除了在個體層級，例如動物的斑紋最為顯著之外，在細胞層級以及分子層級也有自我組織的過程發生。在細胞層級方面，細胞骨骼的自我組織，對於細胞的形態、能動性以及分化都有高度的相關。而在分子層級方面，最被廣泛研究的就是蛋白質的組合結構；而蛋白質異常的結構常會造成人類疾病，例如阿茲海默症。 我們的研究，首先會針對分子層級的蛋白質自我組織現象，以綠螢光修飾肌動蛋白(actin-GFP)作不同酸鹼環境之下的聚集(aggregation)測試，並期許能藉由金奈米粒子特殊的光學特性作為感測。 下一步，則以細胞層級為主要的觀測對象。這部份以C2C12肌肉母細胞(myoblast)在不同的硬度之下，會產生不同的型態作為基礎，研究細胞在不同的幾何限制之下的表現。硬度的調整方面，使用聚丙烯酰酸(polyacrylamide)作為基質材料；而幾何限制則是以微接觸印刷(micro-contact printing)將微纖維連接蛋白(fibronectin)作為接合修飾，以製備出來的基質作細胞培養。觀察的重點則著重在單一細胞內的細胞骨骼的自我組織現象，以及多細胞之間的連接表現。 這個研究可以觀察到不同層級之下的自我組織行為，而這些行為在對於疾病的機轉以及相對應的療癒可能是一個可行的研究途徑。Self-organization is a common phenomenon in the nature world. In biology, in addition to individual level, such as skin patterns, self-organization procedure is generally seen in cellular and molecular levels. In cellular level, self-organization of cytoskeleton is highly connected to cell morphology, motility, and differentiation. In molecular level, protein folding is studied widely. Protein misfolding typically causes human disease such as Alzheimer’s Disease. In our study of molecular scale, self organization of protein was evaluated by observing actin-GFP aggregation in various pH conditions. Moreover, because of the optical characteristics, gold nanoparticle is expected to be used as a sensor. In cellular scale, substrates with various stiffness and geometries were used to evaluate the behaviors of C2C12 myoblast cells. Polyacrylamide hydrogel was used to fabricate substrates with various stiffness. Geometries were controlled using micro-contact printing with fibronectin. Self-organization of cytoskeleton in single patterned cells and connections of multiple cells were observed. In our study, self-organizations in molecular and cellular scales were investigated. This procedure may be an approach to obtain mechanisms of disease and corresponding therapies