The isolation and cultivation of intact, single myofibers presents a superior approach for studying myogenic cells in their native position. The cells’ characteristics remain more similar to muscle tissue than in cell culture. Nevertheless, no routinely used method in higher vertebrates exists. Therefore, we aimed at establishing the isolation and cultivation of single myofibers from porcine muscle. For the first time, we implemented the isolation of intact myofibers from porcine fibularis tertius muscle by enzymatic digestion and their subsequent cultivation under floating conditions. Confocal microscopy showed intact myofibrill structures in isolated myofibers. Myogenic cells were able to proliferate at their parent myofiber as shown by the increase of myonuclear number during culture. Additionally, the described method can be used to investigate myogenic cells migrated from isolated myofibers. These cells expressed myogenic markers and were able to differentiate. In the future, our method can be used for genetic manipulation of cells at myofibers, investigation of growth factors or pharmacological substances, and determination of interactions between myofibers and associated cells. Working with isolated myofibers has the potential to bridge conventional cell culture and animal experiments. Adapting the method to porcine muscle allows for application possibilities in veterinary medicine as well as in biomedical research, which cannot be addressed in rodent model systems.

完整完整的单个肌纤维的分离和培养为研究处于天然位置的成肌细胞提供了一种更好的方法。与细胞培养相比，细胞的特征与肌肉组织的相似性更高。然而，在高等脊椎动物中不存在常规使用的方法。因此，我们旨在从猪肌肉中分离和培养单个肌纤维。我们首次通过酶消化及其在漂浮条件下的培养，从猪腓肠肌分离出完整的肌纤维。共聚焦显微镜检查显示分离的肌纤维中完整的肌原纤维结构。如培养期间肌核数目的增加所表明，肌原细胞能够在其亲本肌纤维上增殖。另外，所述方法可用于研究从分离的肌纤维迁移的成肌细胞。这些细胞表达肌原性标志物并能够分化。将来，我们的方法可用于肌纤维细胞的遗传操作，生长因子或药理物质的研究以及肌纤维与相关细胞之间相互作用的确定。使用隔离的肌纤维可能会桥接常规细胞培养和动物实验。使该方法适应猪肌肉可以在兽医学以及生物医学研究中应用，这在啮齿动物模型系统中无法解决。我们的方法可用于肌纤维细胞的遗传操作，生长因子或药理物质的研究以及肌纤维与相关细胞之间相互作用的确定。使用隔离的肌纤维可能会桥接常规细胞培养和动物实验。使该方法适应猪肌肉可以在兽医学以及生物医学研究中应用，这在啮齿动物模型系统中无法解决。我们的方法可用于肌纤维细胞的遗传操作，生长因子或药理物质的研究以及肌纤维与相关细胞之间相互作用的确定。使用隔离的肌纤维可能会桥接常规细胞培养和动物实验。使该方法适应猪肌肉可以在兽医学以及生物医学研究中应用，这在啮齿动物模型系统中无法解决。