Abstract：
Isolation of both lymphocytes and myogenic cells from muscle tissue is required for elucidating the cellular and molecular mechanisms of muscle regeneration. Here, we aimed to establish an optimal method obtaining a high yield of lymphocytes during muscle regeneration. After the muscle injury, we observed higher infiltration of lymphocytic cells in the muscle on day 3 after injury. Then, we compared two different white blood cell isolation methods, the Percoll gradient and CD45-magnetic bead methods, to assess the percentage and number of T and B cells. Flow cytometry analysis showed that the CD45-magnetic bead method has a better efficiency in isolating CD4+, CD8+ T cells, and B cells from injured muscle tissues of wild-type and mdx mice than that by the Percoll gradient method. Moreover, we found that the CD45-negative fraction from wild-type and mdx mice includes myogenic cells. In conclusion, we report that the CD45-magnetic bead method is suitable to isolate T and B cells during muscle regeneration with higher purity and yield and can also isolate myogenic cells within the same sample. This method provides a technical basis for further studies on muscle regeneration, involving lymphocytes and muscle cells, with a wide range of clinical applications.
摘要：
为了阐明肌肉再生的细胞和分子机制，需要从肌肉组织中分离淋巴细胞和肌源性细胞。这里,我们的目的是建立一种在肌肉再生过程中获得高产量淋巴细胞的最佳方法。肌肉损伤后,我们观察到损伤后第3天肌肉淋巴细胞浸润较高。然后，我们比较了两种不同的白细胞分离方法，Percoll梯度和CD45磁珠方法，评估T细胞和B细胞的百分比和数量。流式细胞仪分析表明,CD45-磁珠法分离CD4+有较好的效果,CD8+T细胞，和B细胞来自野生型和mdx小鼠的毁伤肌肉组织，比照Percoll梯度法。此外，我们发现野生型和mdx小鼠的CD45阴性部分包括肌源性细胞。总之,我们报告说，CD45磁珠法适用于肌肉再生过程中分离T和B细胞，纯度和产量更高，也可以在同一样品中分离成肌细胞。该方法为进一步研究肌肉再生提供了技术基础。涉及淋巴细胞和肌肉细胞，具有广泛的临床应用。
