Department of General Surgery, Yonsei University Wonju College of Medicine, Wonju, Republic of Korea.
Regeneration Medicine Research Center, Yonsei University Wonju College of Medicine, Wonju, Republic of Korea.
Cell Biol Int. 2021 Dec;45(12):2443-2451. doi: 10.1002/cbin.11688. Epub 2021 Aug 23.
Skeletal muscle satellite cells (SkMSCs) play crucial roles in muscle fiber maintenance, repair, and remodeling; however, it remains unknown if these properties are preserved in cultured SkMSCs. In this study, we investigated the characteristics of cultured SkMSCs and their ability to regulate the activity of M1 macrophages. SkMSCs grew well with an average population doubling time of 26.26 ± 6.85 h during 10 passages (P). At P5, Pax7, MyoD, cluster of differentiation (CD)34, and CD56 were not expressed in SkMSCs, but the MSC markers CD73, CD105, and CD90 were expressed and the cells were differentiated into adipocytes and osteoblasts. When SkMSCs were cocultured with macrophages, interleukin (IL)-1β secretion was decreased, prostaglandin (PG)E2 was produced in coculture, and cyclooxygenase-2 protein was induced in an SkMSC-dependent manner. Hepatocyte growth factor (HGF) was highly secreted by monocultured SkMSCs; interferon-γ and lipopolysaccharide reduced its expression level. However, HGF expression recovered when SkMSCs and macrophages were cocultured. Although exogenous PGE2 upregulated macrophage pro-IL-1β expression, it suppressed the secretion of cleaved IL-1β. In contrast, HGF decreased active IL-1β secretion without affecting pro-IL-1β expression. Co-treatment of macrophages with HGF and PGE2 reduced pro-IL-1β expression level and active IL-1β secretion. Our results suggest that SkMSCs lose their satellite cell properties during serial passaging but acquire mesenchymal stem cell properties including the ability to exert an anti-inflammatory response for macrophages through PGE2 and HGF.
骨骼肌卫星细胞(SkMSCs)在肌纤维维持、修复和重塑中发挥着至关重要的作用;然而,其在培养的 SkMSCs 中是否保留这些特性尚不清楚。在本研究中,我们研究了培养的 SkMSCs 的特性及其调节 M1 巨噬细胞活性的能力。SkMSCs 在 10 代培养中生长良好,平均倍增时间为 26.26±6.85 h(P)。在 P5 时,SkMSCs 不表达 Pax7、MyoD、分化群(CD)34 和 CD56,但表达 MSC 标志物 CD73、CD105 和 CD90,并且可以分化为脂肪细胞和成骨细胞。当 SkMSCs 与巨噬细胞共培养时,白细胞介素(IL)-1β 的分泌减少,前列腺素(PG)E2 在共培养中产生,并且环氧化酶-2 蛋白以 SkMSC 依赖的方式被诱导。SkMSCs 单独培养时高度分泌肝细胞生长因子(HGF);干扰素-γ 和脂多糖降低其表达水平。然而,当 SkMSCs 和巨噬细胞共培养时,HGF 的表达恢复。尽管外源性 PGE2 上调了巨噬细胞前 IL-1β 的表达,但它抑制了 cleaved IL-1β 的分泌。相比之下,HGF 降低了活性 IL-1β 的分泌,而不影响前 IL-1β 的表达。巨噬细胞共处理 HGF 和 PGE2 降低了前 IL-1β 的表达水平和活性 IL-1β 的分泌。我们的结果表明,SkMSCs 在连续传代过程中失去了卫星细胞的特性,但获得了间充质干细胞的特性,包括通过 PGE2 和 HGF 对巨噬细胞发挥抗炎反应的能力。