Muscle Biology Laboratory, Department of Animal Sciences, Washington State University, P.O. Box 646310, Pullman, WA, 99164-6310, USA,
Cytotechnology. 2008 Mar;56(3):171-8. doi: 10.1007/s10616-008-9140-5. Epub 2008 Feb 28.
Cell cultures were established from the skeletal muscle tissue of 6-13 months old rainbow trout and 12-14 months old yellow perch. Approximately 27,000 +/- 5,000 cells/g (trout; N = 5) and 5,000 +/- 1,200 cells/g of tissue (perch; N = 4) were obtained. Isolation and propagation were qualitatively greater for both species when the cells (younger fish producer more cells than older fish) were exposed to DMEM + 15% FBS, rather than L-15 + 15% FBS, at 20 degrees C (trout) and at 24 degrees C (yellow perch). Two morphologically distinct cell types were observed in cultures of both species, some of which eventually formed very small myotubes, which displayed immunocytological reactivity for myogenin, myosin heavy chain, and alpha-actinin; the second population of cells remained unstained. Successful cryopreservation was achieved using a 5% DMSO and 95% serum mixture, but post-thawing viabilities were low 5-27% (trout) and 14-30% (perch). Further research is needed in order to determine cell type specificity of isolated cells.
从 6-13 个月大的虹鳟鱼和 12-14 个月大的黄鲈的骨骼肌组织中建立了细胞培养。分别获得了约 27,000 +/- 5,000 个细胞/g（虹鳟鱼；N = 5）和 5,000 +/- 1,200 个细胞/g 的组织（黄鲈；N = 4）。当细胞（幼鱼产生的细胞比老鱼多）在 20 摄氏度（虹鳟鱼）和 24 摄氏度（黄鲈）下暴露于 DMEM + 15% FBS 中时，两种物种的分离和繁殖质量都更高，而不是 L-15 + 15% FBS。在两种物种的培养物中观察到两种形态上明显不同的细胞类型，其中一些最终形成非常小的肌管，这些肌管对肌生成素、肌球蛋白重链和α-肌动蛋白表现出免疫细胞化学反应；第二群细胞保持未染色。使用 5% DMSO 和 95%血清混合物成功实现了冷冻保存，但解冻后的存活率较低，为 5-27%（虹鳟鱼）和 14-30%（黄鲈）。为了确定分离细胞的细胞类型特异性，还需要进一步研究。
