Cytoskeletal-Mediated Tension Modulates the Directed Self-Assembly of Microtissues
Cytoskeletal-Mediated Tension Modulates the Directed Self-Assembly of Microtissues
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DOI：
10.1089/ten.tea.2007.0320
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发表时间：
2008-12-01
影响因子：
4.1
通讯作者：
Morgan, Jeffrey R.
中科院分区：
文献类型：
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作者：
Dean, Dylan M.;Morgan, Jeffrey R.
细胞自组装成微组织（如球体）的能力主要归因于通过结合表面膜蛋白（如钙粘蛋白）实现的细胞间凝聚力。然而，高度动态和复杂的细胞骨架重排与这些结合事件相协调，因此可能参与自组装。通过抑制这种细胞骨架活性，Y-27632已被用于预防和治疗纤维化疾病。在这里，我们使用Rho激酶抑制剂来研究细胞收缩在自组装中的作用。正常人成纤维细胞（NHF），H35肝癌细胞（H35），和两者的混合物（杂交）在非粘性，微模塑水凝胶中的微组织的定向自组装过程中用药物处理。受约束和不受约束的NHF微组织的自组装动力学被药物显着减慢，并且抑制是剂量响应性和可逆的。尽管NHF和H35的分选在药物存在下正常发生，但当与未处理的NHF混合时，Y-27632处理的NHF分选到球状体的外部。当在槽微模具中与H35混合时，NHF可以驱动从杂交微组织的核心形成球状体，即使相对于H35（1：19）数量较少。这些发现表明，细胞收缩控制自组装的动力学，并表明微组织核心内的NHF可以通过与H35的异型键传递收缩力。使用成纤维细胞和收缩抑制剂控制定向自组装可用于体外组织工程以及代表纤维化疾病的体外模型。
The ability of cells to self-assemble into a microtissue such as a spheroid has been attributed mainly to intercellular cohesiveness achieved by the binding of surface membrane proteins such as cadherins. However, highly dynamic and complex cytoskeletal rearrangements are coordinated with these binding events and therefore are likely to participate in self-assembly. By inhibiting such cytoskeletal activity, Y-27632 has been used to prevent and treat fibrotic disease. Here, we used the Rho kinase inhibitor to investigate the role that cellular contraction plays in self-assembly. Normal human fibroblasts (NHFs), Reuber-H35 hepatoma cells (H35s), and mixes of the two (hybrid) were treated with drug during directed self-assembly of microtissues in nonadhesive, micromolded hydrogels. The kinetics of self-assembly of both constrained and unconstrained NHF microtissues were dramatically slowed by the drug, and inhibition was dose responsive and reversible. Although sorting of NHFs and H35s occurred normally in the presence of drug, Y-27632-treated NHFs sorted to the outside of a spheroid when mixed with untreated NHFs. When mixed with H35s in trough micromolds, NHFs could drive spheroid formation from the core of the hybrid microtissues even in small numbers relative to H35s (1:19). These findings demonstrate that cellular contraction controls the kinetics of self-assembly and suggests that NHFs within the core of a microtissue can transmit contractile forces through heterotypic bonds with H35s. The control of directed self-assembly using fibroblasts and contraction inhibitors may be useful for in vitro tissue engineering as well as represent an in vitro model for fibrotic disease.