Collagen Microparticle-Mediated 3D Cell Organization: A Facile Route to Bottom-up Engineering of Thick and Porous Tissues.
Collagen Microparticle-Mediated 3D Cell Organization: A Facile Route to Bottom-up Engineering of Thick and Porous Tissues.
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DOI：
10.1021/acsbiomaterials.7b00131
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发表时间：
2017-08
影响因子：
5.8
通讯作者：
Yuya Yajima;M. Yamada;R. Utoh;M. Seki
中科院分区：
文献类型：
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作者：
Yuya Yajima;M. Yamada;R. Utoh;M. Seki
在紧密堆积的人工3D细胞结构中，由于氧气和营养的供应有限，位于结构中心附近的细胞不能发挥功能。在这里，我们描述了一种简单、独特和高度通用的方法，将细胞大小的胶原蛋白微粒作为颗粒支架，将细胞组织成厚实但多孔的3D组织。当细胞和颗粒混合并种植在非细胞黏附的平面室中时，它们聚集在一起形成厚度为100-150μm的片状结构。在该结构中，均匀分布的颗粒作为细胞之间的粘合剂，调节细胞间的强烈收缩。我们证实了几个因素，包括颗粒/细胞比和颗粒大小，对用小鼠胚胎成纤维细胞(NIH-3T3细胞)制备的多孔组织的稳定性和收缩行为有重要影响。横切面观察以及细胞增殖和活性分析表明，组成组织的细胞具有功能，主要是因为细胞/颗粒之间的内部毛孔作为有效的分子运输途径。此外，我们利用人肝癌细胞(HepG2细胞)制备了肝脏模型的厚细胞组织，并证实了当用几种不同的稳定方案(戊二醛、京尼平和乙酸甲酯)制备的胶原微粒形成复合组织时，肝脏特异性功能被上调。所提出的方法对于一步生产孔隙率和形态可调的厚孔结构是非常有用的。
In closely packed artificial 3D cellular constructs, cells located near the center of the constructs are not functional because of the limited supply of oxygen and nutrition. Here we describe a simple, unique, and highly versatile approach to organizing cells into thick but porous 3D tissues, using cell-sized collagen microparticles as particulate scaffolds. When cells and particles are mixed and seeded in a noncell-adhesive planar chamber, they gather to form sheet-shaped structures with a thickness of 100-150 μm. In the construct, uniformly distributed particles work as a binder between cells and modulate the strong intercellular contraction. We confirmed that several factors, including the particle/cell ratio and particle size, critically affect the stability and shrink behaviors of porous tissues prepared using mouse embryonic fibroblasts (NIH-3T3 cells). Cross-sectional observation, together with cell proliferation and viability assays, revealed that the cells composing the tissues are functional primarily because interior pores between cells/particles worked as a path for efficient molecular transport. Furthermore, we prepared thick cell tissues of a liver model using human hepatocarcinoma cells (HepG2 cells), and confirmed that liver-specific functions were upregulated when composite tissues were formed using collagen microparticles prepared with several different stabilization protocols by glutaraldehyde, genipin, and methyl acetate). The process presented would be highly useful in enabling one-step production of thick cellular constructs in which porosity and morphology are tunable.