介質硬度及水壓對肌母細胞型態之影響

Abstract

細胞移動，是多細胞生物組織在許多生理及及病理過程，包括組織發育、組織維持、血管新生、發炎、老化、傷口癒合、再生以及腫瘤發生和腫瘤轉移中極重要的步驟，而細胞的型態又與細胞發生移動前息息相關，故我們將聚焦在給予不同的力學微環境下細胞型態的變化。 本實驗的目的是藉由設計出一可調硬度的生物材料微環境系統，觀察C2C12肌母細胞在不同硬度的微環境和外加靜態水壓下對其細胞型態的影響。而我們使用的是PAAM水膠，PAAM水膠為生物力學領域中被相當廣泛應用的一種生物材料，因其方便性，藉由調整acrylamide:bisacrylamide的濃度比例，便可使其硬度範圍落在目標區間，以本實驗的設計來說為3kPa~40kPa。 在先前的實驗和本實驗中都觀察到相同的結論，也就是將C2C12肌母細胞種植在較軟的水膠微環境下，會得到較小及較圓的細胞型態分佈，而隨著微環境硬度增加，其細胞型態也由之前圓型分佈，逐漸轉為拉長的纖維狀的型態分佈。另外，在實行24小時靜態水壓後，不論是在何種硬度的微環境上，都有觀察到較沒有施行水壓的細胞更為明顯的伸長纖維型態。

Cell migration is a hugely important step in the physiological and pathological processes of multicellular organisms, such as tissue development, tissue maintenance, angiogenesis, inflammation, aging, wound healing or regeneration, tumorigenesis, metastasis, etc. and often plays a crucial role in determining morphogenesis. The purpose of this project was to design and characterize a biomaterial system with a range of substrate stiffness for the use of biological experiments investigating the effect of substrate stiffness on the morphology and the effect of C2C12 cells exposuring in a hydrostatic pressure environment. Polyacrylamide (PAAM) was used as the biomaterial. This is a hydrogel with tunable stiffness through chemical cross-linking. Modulus of elasticity with PAAM of a range of acrylamide:bis-acrylamide concentrations yielded a stiffness range of 3kPa to 40kPa. Initial cellular experiments showed that C2C12 seeded on softer substrates were smaller and round and progressively became more elongated and fibroblastic as the stiffness of the substrate was increased.