Disulfide-Based Diblock Copolymer Worm Gels: A Wholly-Synthetic Thermoreversible 3D Matrix for Sheet-Based Cultures.
Disulfide-Based Diblock Copolymer Worm Gels: A Wholly-Synthetic Thermoreversible 3D Matrix for Sheet-Based Cultures.
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DOI：
10.1021/acs.biomac.5b01266
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发表时间：
2015-11
影响因子：
6.2
通讯作者：
Karen A. Simon;N. Warren;B. Mosadegh;M. R. Mohammady;G. Whitesides;S. Armes
中科院分区：
文献类型：
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作者：
Karen A. Simon;N. Warren;B. Mosadegh;M. R. Mohammady;G. Whitesides;S. Armes
众所周知，3D体外细胞培养提供了比2D细胞培养更好的模型，用于理解细胞的体内微环境。然而，在处理和分析3D细胞培养物方面仍然存在重大的技术挑战，这限制了它们的广泛应用。在此，我们展示了完全合成的温敏嵌段共聚物蠕虫在基于片材的3D细胞培养中的应用。这些蠕虫在20-37 °C下可逆地形成柔软的独立式凝胶，在冷却至5 °C时可以迅速转化为自由流动的球体分散体。发现具有二硫化物基团的蠕虫的官能化对于确保这些水凝胶的足够机械稳定性以实现长期细胞培养是必不可少的。这些二硫化物基团通过基于二硫化物的二甲基丙烯酸酯在有利于分子内环化的条件下的统计共聚方便地引入，并且随后的硫醇/二硫化物交换导致凝胶内相邻蠕虫之间形成可逆的共价键。这种新方法使细胞能够包埋在具有良好活力的微米厚凝胶板中，允许细胞培养至少12天，并只需在4 °C的缓冲液中孵育培养物即可促进从凝胶中回收活细胞（因此，避免了使用商业蛋白质凝胶（例如Matrigel）时细胞收获所需的酶降解）。
It is well-known that 3D in vitro cell cultures provide a much better model than 2D cell cultures for understanding the in vivo microenvironment of cells. However, significant technical challenges in handling and analyzing 3D cell cultures remain, which currently limits their widespread application. Herein, we demonstrate the application of wholly synthetic thermoresponsive block copolymer worms in sheet-based 3D cell culture. These worms form a soft, free-standing gel reversibly at 20-37 °C, which can be rapidly converted into a free-flowing dispersion of spheres on cooling to 5 °C. Functionalization of the worms with disulfide groups was found to be essential for ensuring sufficient mechanical stability of these hydrogels to enable long-term cell culture. These disulfide groups are conveniently introduced via statistical copolymerization of a disulfide-based dimethacrylate under conditions that favor intramolecular cyclization and subsequent thiol/disulfide exchange leads to the formation of reversible covalent bonds between adjacent worms within the gel. This new approach enables cells to be embedded within micrometer-thick slabs of gel with good viability, permits cell culture for at least 12 days, and facilitates recovery of viable cells from the gel simply by incubating the culture in buffer at 4 °C (thus, avoiding the enzymatic degradation required for cell harvesting when using commercial protein-based gels, such as Matrigel).