Micropatterning Alginate Substrates for in vitro Cardiovascular Muscle on a Chip.
Micropatterning Alginate Substrates for in vitro Cardiovascular Muscle on a Chip.
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DOI：
10.1002/adfm.201203319
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发表时间：
2013-08-12
影响因子：
19
通讯作者：
Parker KK
中科院分区：
文献类型：
--
作者：
Agarwal A;Farouz Y;Nesmith AP;Deravi LF;McCain ML;Parker KK
软水凝胶如藻酸盐是体外构建肌肉的理想基质，因为它们具有接近体内细胞外基质（ECM）网络的结构和机械性质。然而，水凝胶通常不适于蛋白质粘附和图案化。此外，肌肉结构和其基础ECM是高度各向异性的，并且由于这些系统中结构和功能之间的紧密耦合，体外模型必须概括重建组织中的结构各向异性以用于体内相关性。我们提出了两种技术，以创建化学和结构的异质性软藻酸盐基板内，并利用它们来工程各向异性肌肉单层：（i）微接触印刷线的细胞外基质蛋白质的平坦藻酸盐基板上，以引导细胞过程与化学线索，和（ii）微模塑的藻酸盐表面成凹槽和脊，以引导细胞过程与地形线索。新生大鼠心室肌细胞以及人脐动脉血管平滑肌细胞成功地附着到这两个微图案基板，导致随后形成的各向异性横纹和平滑肌组织。从这些构建体上切下的肌肉薄膜支架然后用于工程化肌肉组织的功能表征。因此，微图案化的藻酸盐是肌肉组织体外模型的理想基质，因为它有助于再现肌肉的各向异性结构，模拟ECM微环境的机械特性，并且适合于功能收缩特性的评价。
Soft hydrogels such as alginate are ideal substrates for building muscle in vitro because they have structural and mechanical properties close to the in vivo extracellular matrix (ECM) network. However, hydrogels are generally not amenable to protein adhesion and patterning. Moreover, muscle structures and their underlying ECM are highly anisotropic, and it is imperative that in vitro models recapitulate the structural anisotropy in reconstructed tissues for in vivo relevance due to the tight coupling between sturcture and function in these systems. We present two techniques to create chemical and structural heterogeneities within soft alginate substrates and employ them to engineer anisotropic muscle monolayers: (i) microcontact printing lines of extracellular matrix proteins on flat alginate substrates to guide cellular processes with chemical cues, and (ii) micromolding of alginate surface into grooves and ridges to guide cellular processes with topographical cues. Neonatal rat ventricular myocytes as well as human umbilical artery vascular smooth muscle cells successfully attach to both these micropatterned substrates leading to subsequent formation of anisotropic striated and smooth muscle tissues. Muscular thin film cantilevers cut from these constructs are then employed for functional characterization of engineered muscular tissues. Thus, micropatterned alginate is an ideal substrate for in vitro models of muscle tissue because it facilitates recapitulation of the anisotropic architecture of muscle, mimics the mechanical properties of the ECM microenvironment, and is amenable to evaluation of functional contractile properties.