纤维多异氰化物水凝胶用于3D细胞培养应用。

IF 13.1 1区 生物学 Q1 BIOCHEMICAL RESEARCH METHODS
Hongbo Yuan, Kaizheng Liu, Melissa J J van Velthoven, Jyoti Kumari, Yuying Bao, Susana Rocha, Paul H J Kouwer
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引用次数: 0

摘要

基于水凝胶的三维(3D)细胞培养模型正迅速发展成为组织工程、机械生物学、疾病建模和药物筛选的重要工具。虽然近年来出现了各种各样的合成凝胶,但它们未能充分渗透市场,主要有两个原因:它们难以模拟细胞外基质,或者难以用于凝胶形成和细胞提取。模拟自然界的复杂性是具有挑战性的:细胞外基质在细胞发育和功能中起着至关重要的作用,这远远超出了简单的机械支持。最近,我们推出了用于3D细胞培养应用的聚异氰酸酯(PIC)水凝胶。纤维结构和相关的(非)线性力学行为与生物凝胶(如胶原蛋白和纤维蛋白)的物理特性非常相似。作为全合成材料,PIC凝胶具有高度的可定制性和可重复性。此外,PIC凝胶的热响应特性使其易于在实验室中处理;凝胶在37°C下立即形成,细胞在冷却到5°C后很容易提取。PIC凝胶的潜力已经在讨论不同细胞系、原代细胞和类器官以及体内实验的快速扩展的论文库中得到证明。这份手稿提供了如何处理PIC凝胶在化学和细胞生物学实验室的协议。材料制备需要72小时,细胞封装需要1小时,下游分析的时间取决于所使用的(商业)方法。所描述的方案适用于具有细胞培养和分子生物学专业知识的研究人员。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Fibrous polyisocyanide hydrogels for 3D cell culture applications.

Three-dimensional (3D) cell culture models based on hydrogels are rapidly evolving into a prominent tool for tissue engineering, mechanobiology, disease modeling and drug screening. While a vast variety of synthetic gels have emerged in recent years, they fail to penetrate the market substantially for two major reasons: they poorly mimic the extracellular matrix or they are difficult to use in gel formation and cell extraction. Mimicking the complexity of nature is challenging: the extracellular matrix plays a crucial role in cell development and function, which goes well beyond simple mechanical support. Recently, we introduced polyisocyanide (PIC) hydrogels for 3D cell culture applications. The fibrous architecture and associated (non)linear mechanical behavior closely mimic the physical properties of biogels such as collagen and fibrin. As fully synthetic materials, PIC gels benefit from high tailorability and reproducibility. Moreover, the thermoresponsive properties of PIC gels make them easy to handle in the lab; the gels form instantly at 37 °C and cells are easily extracted after cooling to 5 °C. The potential of PIC gels has been demonstrated in a quickly expanding library of papers discussing different cell lines, primary cells and organoids, as well as in vivo experiments. This manuscript provides protocols on how to handle PIC gels in the chemistry and cell biology laboratories. Material preparation requires 72 h. Cell encapsulation takes 1 h and the time for downstream analysis depends on the (commercial) methods used. The protocols described are suitable for researchers with expertise in cell culture and molecular biology.

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来源期刊
Nature Protocols
Nature Protocols 生物-生化研究方法
CiteScore
29.10
自引率
0.70%
发文量
128
审稿时长
4 months
期刊介绍: Nature Protocols focuses on publishing protocols used to address significant biological and biomedical science research questions, including methods grounded in physics and chemistry with practical applications to biological problems. The journal caters to a primary audience of research scientists and, as such, exclusively publishes protocols with research applications. Protocols primarily aimed at influencing patient management and treatment decisions are not featured. The specific techniques covered encompass a wide range, including but not limited to: Biochemistry, Cell biology, Cell culture, Chemical modification, Computational biology, Developmental biology, Epigenomics, Genetic analysis, Genetic modification, Genomics, Imaging, Immunology, Isolation, purification, and separation, Lipidomics, Metabolomics, Microbiology, Model organisms, Nanotechnology, Neuroscience, Nucleic-acid-based molecular biology, Pharmacology, Plant biology, Protein analysis, Proteomics, Spectroscopy, Structural biology, Synthetic chemistry, Tissue culture, Toxicology, and Virology.
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