Principles of Hanging Drop Method (Spheroid Formation) in Cell Culture.

Q4 Biochemistry, Genetics and Molecular Biology Methods in molecular biology Pub Date : 2024-02-28 DOI:10.1007/7651_2024_527
Mohammad Rasouli, Fatemeh Safari, Mohammad Hossein Kanani, Hiva Ahvati
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Abstract

A type of three-dimensional (3D) cell culture models which is simple and easy is hanging drop method. The hanging drop method emerges as a pivotal technique with diverse applications in cancer research and cell biology. This method facilitates the formation of multicellular spheroids, providing a unique environment for studying cell behavior dynamics. The hanging drop method's theoretical underpinning relies on gravity-enforced self-assembly, allowing for cost-effective, reproducible 3D cell cultures with controlled spheroid sizes. The advantages of this approach include its efficiency in producing cellular heterogeneity, particularly in non-adherent 3D cultures, and its ability to create hypoxic spheroids, making it a suitable model for studying cancer. Moreover, the hanging drop method has proven valuable in investigating various aspects such as tissue structure, signaling pathways, immune activation of cancer cells, and notably, cell proliferation. Researchers have utilized the hanging drop method to explore the dynamics of cell proliferation, studying the effects of mesenchymal stem cells (MSC) secretome on cancer cells. The method's application involves co-culturing different cell lines, assessing spheroid formations, and quantifying their sizes over time. These studies have unveiled intricate cell behavior dynamics, demonstrating how the MSC secretome influences cancer cell growth and viability within a three-dimensional co-culture paradigm.

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细胞培养中的悬滴法(球体形成)原理。
悬滴法是一种简单易行的三维(3D)细胞培养模型。悬滴法是一种关键技术,在癌症研究和细胞生物学中有着广泛的应用。这种方法有助于形成多细胞球体,为研究细胞行为动态提供了独特的环境。悬滴法的理论基础依赖于重力强化自组装,可实现成本效益高、可重复的三维细胞培养,球体大小可控。这种方法的优点包括能有效产生细胞异质性,特别是在非粘附三维培养物中,还能产生缺氧球体,因此适合作为研究癌症的模型。此外,悬滴法在研究组织结构、信号通路、癌细胞的免疫激活等各方面都很有价值,尤其是在细胞增殖方面。研究人员利用悬滴法探索细胞增殖动态,研究间充质干细胞(MSC)分泌物对癌细胞的影响。该方法的应用包括共同培养不同的细胞系,评估球形细胞的形成,并随时间变化量化其大小。这些研究揭示了错综复杂的细胞行为动态,展示了间充质干细胞分泌组如何在三维共培养范例中影响癌细胞的生长和存活率。
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来源期刊
Methods in molecular biology
Methods in molecular biology Biochemistry, Genetics and Molecular Biology-Genetics
CiteScore
2.00
自引率
0.00%
发文量
3536
期刊介绍: For over 20 years, biological scientists have come to rely on the research protocols and methodologies in the critically acclaimed Methods in Molecular Biology series. The series was the first to introduce the step-by-step protocols approach that has become the standard in all biomedical protocol publishing. Each protocol is provided in readily-reproducible step-by-step fashion, opening with an introductory overview, a list of the materials and reagents needed to complete the experiment, and followed by a detailed procedure that is supported with a helpful notes section offering tips and tricks of the trade as well as troubleshooting advice.
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