Department of Medicine, Center for Biomedical Engineering, Brigham and Women's Hospital, Harvard Medical School, 65 Landsdowne Street, Rm 265, Cambridge, Massachusetts 02139, USA.
Biotechnol Bioeng. 2010 May 1;106(1):138-48. doi: 10.1002/bit.22667.
In this article, we describe an approach to generate microporous cell-laden hydrogels for fabricating biomimetic tissue engineered constructs. Micropores at different length scales were fabricated in cell-laden hydrogels by micromolding fluidic channels and leaching sucrose crystals. Microengineered channels were created within cell-laden hydrogel precursors containing agarose solution mixed with sucrose crystals. The rapid cooling of the agarose solution was used to gel the solution and form micropores in place of the sucrose crystals. The sucrose leaching process generated homogeneously distributed micropores within the gels, while enabling the direct immobilization of cells within the gels. We also characterized the physical, mechanical, and biological properties (i.e., microporosity, diffusivity, and cell viability) of cell-laden agarose gels as a function of engineered porosity. The microporosity was controlled from 0% to 40% and the diffusivity of molecules in the porous agarose gels increased as compared to controls. Furthermore, the viability of human hepatic carcinoma cells that were cultured in microporous agarose gels corresponded to the diffusion profile generated away from the microchannels. Based on their enhanced diffusive properties, microporous cell-laden hydrogels containing a microengineered fluidic channel can be a useful tool for generating tissue structures for regenerative medicine and drug discovery applications.
在本文中,我们描述了一种生成用于制造仿生组织工程构建体的微孔细胞负载水凝胶的方法。通过微成型流道和浸出蔗糖晶体,在细胞负载水凝胶中制造出不同长度尺度的微孔。在含有琼脂糖溶液和蔗糖晶体的细胞负载水凝胶前体中创建了微工程通道。琼脂糖溶液的快速冷却用于使溶液凝胶化并在蔗糖晶体的位置形成微孔。蔗糖浸出过程在凝胶中产生了均匀分布的微孔,同时能够将细胞直接固定在凝胶中。我们还表征了作为工程孔隙率函数的细胞负载琼脂糖凝胶的物理、机械和生物学特性(即微孔率、扩散率和细胞活力)。微孔率可控制在 0%至 40%之间,多孔琼脂糖凝胶中分子的扩散率与对照相比有所增加。此外,在微孔琼脂糖凝胶中培养的人肝癌细胞的活力与远离微通道生成的扩散曲线相对应。基于其增强的扩散性能,含有微工程流道的微孔细胞负载水凝胶可以成为用于再生医学和药物发现应用的组织结构生成的有用工具。