Fusion of Human Synovium-Derived Mesenchymal Stem/Stromal Cells with Primary Human Chondrocytes Using the Modified Adherence Method (MAM).

Q4 Biochemistry, Genetics and Molecular Biology Methods in molecular biology Pub Date : 2025-01-01 DOI:10.1007/7651_2025_620
Tilen Burnik, Janja Zupan, Matjaž Jeras, Maša Kandušer
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Abstract

Cell fusion is a complex phenomenon that is key in maintaining tissue homeostasis, particularly in aiding tissue regeneration processes. Studies show that mesenchymal stem/stromal cells (MSCs) are capable of restoring damaged tissue by adopting the phenotype of various cell types via cell fusion. As cell fusion of MSCs with cells of different origin remains poorly researched, we have developed a protocol that allows successful electrofusion between human synovium-derived MSCs and human chondrocytes. Building on from our protocol can help researchers study the cell fusion processes in the in vitro environment and could set basis for development of fusion cell-based advanced therapy medicinal products (ATMPs). In our protocol, we provide a detailed description on how to culture both of the fusion partner cells, how to carry out the modified adherence method (MAM) to achieve a high yield of successfully fused cells, and how to determine the yield of cell fusion using either methyl violet or fluorescent cell trackers.

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改良黏附法(MAM)融合人滑膜源间充质干细胞/基质细胞与原代人软骨细胞。
细胞融合是一种复杂的现象,是维持组织稳态的关键,特别是在帮助组织再生过程中。研究表明,间充质干细胞(mesenchymal stem/stromal cells, MSCs)通过细胞融合,采用多种细胞类型的表型,具有修复受损组织的能力。由于间充质干细胞与不同来源细胞的细胞融合研究仍然很少,我们已经开发了一种方案,允许人滑膜来源的间充质干细胞与人软骨细胞成功电融合。基于我们的方案可以帮助研究人员在体外环境中研究细胞融合过程,并为融合细胞为基础的先进治疗药物(ATMPs)的开发奠定基础。在我们的方案中,我们详细描述了如何培养融合伴侣细胞,如何实施改进的粘附方法(MAM)以实现高产量的成功融合细胞,以及如何使用甲基紫或荧光细胞跟踪器确定细胞融合的产量。
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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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