Burnik Tilen, Zupan Janja, Jeras Matjaž, Kandušer Maša
Department of Clinical Biochemistry, Faculty of Pharmacy, University of Ljubljana, Ljubljana, Slovenia.
Institute of Pharmacy, Faculty of Pharmacy, University of Ljubljana, Ljubljana, Slovenia.
Methods Mol Biol. 2025;2960:27-37. doi: 10.1007/7651_2025_620.
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.
细胞融合是一种复杂的现象，对于维持组织稳态至关重要，尤其是在协助组织再生过程中。研究表明，间充质干/基质细胞（MSCs）能够通过细胞融合采用各种细胞类型的表型来修复受损组织。由于MSCs与不同来源细胞的细胞融合研究仍较少，我们开发了一种方案，可实现人滑膜来源的MSCs与人类软骨细胞之间的成功电融合。基于我们的方案开展研究，有助于研究人员在体外环境中研究细胞融合过程，并可为基于融合细胞的先进治疗药品（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两种融合伙伴细胞，如何进行改良贴壁法（MAM）以获得高产率的成功融合细胞，以及如何使用甲基紫或荧光细胞追踪剂确定细胞融合率。
