Institute of Fisheries Sciences, Pukyong National University, Gijang-gun, Busan 46041, Republic of Korea.
Division of Fisheries Life Sciences, Pukyong National University, Nam-gu, Busan 48513, Republic of Korea.
Cells. 2023 Sep 21;12(18):2325. doi: 10.3390/cells12182325.
Olive flounder () muscle satellite cells (OFMCs) were obtained by enzymatic primary cell isolation and the explant method. Enzymatic isolation yielded cells that reached 80% confluence within 8 days, compared to 15 days for the explant method. Optimal OFMC growth was observed in 20% fetal bovine serum at 28 °C with 0.8 mM CaCl and the basic fibroblast growth factor (BFGF) to enhance cell growth. OFMCs have become permanent cell lines through the spontaneous immortalization crisis at the 20th passage. Olive flounder skeletal muscle myoblasts were induced into a mitogen-poor medium containing 2% horse serum for differentiation; they fused to form multinucleate myotubes. The results indicated complete differentiation of myoblasts into myotubes; we also detected the expression of the myogenic regulatory factors myoD, myogenin, and desmin. Upregulation (Myogenin, desmin) and downregulation (MyoD) of muscle regulation factors confirmed the differentiation in OFMCs.
牙鲆（）肌肉卫星细胞（OFMCs）通过酶解原代细胞分离和组织块法获得。与组织块法相比，酶解法获得的细胞在 8 天内达到 80%的汇合度，而组织块法需要 15 天。在 28°C 下，最佳的 OFMC 生长条件为 20%胎牛血清、0.8 mM CaCl 和碱性成纤维细胞生长因子（BFGF），以促进细胞生长。OFMCs 在第 20 代时通过自发的永生化危机成为永久细胞系。牙鲆骨骼肌成肌细胞被诱导进入含有 2%马血清的低有丝分裂培养基中进行分化；它们融合形成多核肌管。结果表明成肌细胞完全分化为肌管；我们还检测到肌源性调节因子 MyoD、myogenin 和 desmin 的表达。肌肉调节因子的上调（Myogenin、desmin）和下调（MyoD）证实了 OFMCs 的分化。
