Biomedical Engineering Department, Tissue Engineering Resource Center, Tufts University, 4 Colby St, Medford, MA, 02155, USA.
Research Technology, Tufts University, 16 Dearborn Rd, Somerville, MA, 02144, USA.
Sci Rep. 2023 Mar 29;13(1):5098. doi: 10.1038/s41598-023-31822-2.
Cell-cultivated fish offers the potential for a more ethical, sustainable, and safe seafood system. However, fish cell culture is relatively understudied in comparison to mammalian cells. Here, we established and characterized a continuous Atlantic mackerel (Scomber scombrus) skeletal muscle cell line ("Mack" cells). The cells were isolated from muscle biopsies of fresh-caught fish, with separate isolations performed from two distinct fish. Mack1 cells (cells from the first isolation) were cultured for over a year and subcultured over 130 times. The cells proliferated at initial doubling times of 63.9 h (± 19.1 SD). After a spontaneous immortalization crisis from passages 37-43, the cells proliferated at doubling times of 24.3 h (± 4.91 SD). A muscle phenotype was confirmed through characterization of muscle stemness and differentiation via paired-box protein 7 and myosin heavy chain immunostaining, respectively. An adipocyte-like phenotype was also demonstrated for the cells through lipid accumulation, confirmed via Oil Red O staining and quantification of neutral lipids. New qPCR primers (HPRT, PAX3B, MYOD1, MYOG, TNNT3A, and PPARG) were tailored to the mackerel genome and used to characterize mackerel cell genotypes. This work provides the first spontaneously immortalized fish muscle cell line for research, ideally serving as a reference for subsequent investigation.
细胞培养的鱼类为更具伦理道德、可持续和安全的海鲜系统提供了潜力。然而，与哺乳动物细胞相比，鱼类细胞培养的研究相对较少。在这里，我们建立并鉴定了一种连续的大西洋鲭鱼（Scomber scombrus）骨骼肌细胞系（“Mack”细胞）。这些细胞是从新鲜捕获的鱼的肌肉活检中分离出来的，分别从两条不同的鱼中进行了分离。Mack1 细胞（来自第一次分离的细胞）培养了一年多，并进行了超过 130 次传代培养。细胞最初的倍增时间为 63.9 小时（±19.1 SD）。在第 37-43 代自发永生化危机后，细胞的倍增时间为 24.3 小时（±4.91 SD）。通过配对盒蛋白 7 和肌球蛋白重链免疫染色分别对肌肉干性和分化进行特征鉴定，证实了肌肉表型。通过油红 O 染色和中性脂质定量，证实了细胞中也存在脂肪细胞样表型。针对鲭鱼基因组定制了新的 qPCR 引物（HPRT、PAX3B、MYOD1、MYOG、TNNT3A 和 PPARG），用于鉴定鲭鱼细胞基因型。这项工作提供了第一个用于研究的自发永生化鱼类肌肉细胞系，理想情况下可作为后续研究的参考。
