Kornmuller Anna, Brown Cody F C, Yu Claire, Flynn Lauren E
Biomedical Engineering Graduate Program, The University of Western Ontario.
Department of Anatomy & Cell Biology, Schulich School of Medicine & Dentistry, The University of Western Ontario.
J Vis Exp. 2017 Apr 11(122):55436. doi: 10.3791/55436.
Cell function is mediated by interactions with the extracellular matrix (ECM), which has complex tissue-specific composition and architecture. The focus of this article is on the methods for fabricating ECM-derived porous foams and microcarriers for use as biologically-relevant substrates in advanced 3D in vitro cell culture models or as pro-regenerative scaffolds and cell delivery systems for tissue engineering and regenerative medicine. Using decellularized tissues or purified insoluble collagen as a starting material, the techniques can be applied to synthesize a broad array of tissue-specific bioscaffolds with customizable geometries. The approach involves mechanical processing and mild enzymatic digestion to yield an ECM suspension that is used to fabricate the three-dimensional foams or microcarriers through controlled freezing and lyophilization procedures. These pure ECM-derived scaffolds are highly porous, yet stable without the need for chemical crosslinking agents or other additives that may negatively impact cell function. The scaffold properties can be tuned to some extent by varying factors such as the ECM suspension concentration, mechanical processing methods, or synthesis conditions. In general, the scaffolds are robust and easy to handle, and can be processed as tissues for most standard biological assays, providing a versatile and user-friendly 3D cell culture platform that mimics the native ECM composition. Overall, these straightforward methods for fabricating customized ECM-derived foams and microcarriers may be of interest to both biologists and biomedical engineers as tissue-specific cell-instructive platforms for in vitro and in vivo applications.
细胞功能是通过与细胞外基质(ECM)的相互作用来介导的,细胞外基质具有复杂的组织特异性组成和结构。本文的重点是制造源自ECM的多孔泡沫和微载体的方法,这些泡沫和微载体可用作先进的3D体外细胞培养模型中与生物学相关的基质,或用作组织工程和再生医学的促再生支架及细胞递送系统。以脱细胞组织或纯化的不溶性胶原蛋白为起始材料,这些技术可用于合成具有可定制几何形状的多种组织特异性生物支架。该方法包括机械加工和温和的酶消化,以产生ECM悬浮液,通过控制冷冻和冻干程序,该悬浮液用于制造三维泡沫或微载体。这些源自纯ECM的支架具有高度多孔性,但无需化学交联剂或其他可能对细胞功能产生负面影响的添加剂即可保持稳定。通过改变诸如ECM悬浮液浓度、机械加工方法或合成条件等因素,可在一定程度上调整支架性能。一般来说,这些支架坚固且易于操作,并且可以作为组织进行大多数标准生物学检测的处理,提供了一个模拟天然ECM组成的通用且用户友好的3D细胞培养平台。总体而言,这些制造定制的源自ECM的泡沫和微载体的直接方法,可能会引起生物学家和生物医学工程师的兴趣,作为用于体外和体内应用的组织特异性细胞指导平台。