Key Laboratory of Healthy Mariculture for the East China Sea, Ministry of Agriculture and Rural Affairs, Fisheries College, Jimei University, Xiamen, 361021, China.
College of Ocean and Earth Sciences, Xiamen University, Xiamen, 361005, China.
Mar Biotechnol (NY). 2024 Dec;26(6):1287-1306. doi: 10.1007/s10126-024-10372-2. Epub 2024 Sep 9.
Induced pluripotent stem cells (iPSCs) are a new type of pluripotent cells reprogrammed from somatic cells back into an embryonic-like pluripotent state of stem cells to study development, disease and potential gene therapies. The induction and regulation mechanisms of iPSCs in fish are still unclear. By using the transfection technique, we investigated the crucial function of the OSKMNL factor co-expression for somatic reprogramming in the muscle cell line of large yellow croaker (Larimichthys crocea) (LYCMs) and successfully established a stable iPSCs line (Lc-OSNL-iPSCs). Stable culturing of iPSCs with high alkaline phosphatase activity and a stable karyotype was achieved. The qRT-PCR and immunofluorescence labeling results revealed that Lc-OSNL-iPSCs displayed a high expression level of pluripotent marker genes such as Nanog, Oct4, and Sox2. There were significant differences between Lc-OSNL-iPSCs, Lc-OSKMNL-iPSCs, and LYCMs, and the expression of several genes in maintaining cell pluripotency was up-regulated when the pluripotency signal pathway of stem cells was activated. The technical system for inducing iPSCs of Larimichthys crocea was constructed in this study. This system can serve as a basic model to understand germ cell differentiation mechanism, gender control, genetics, and breeding of large yellow croaker and a platform for studying iPSCs in fish. Interestingly, the acquired iPSCs serves as a useful material for the directional induction of muscle stem cells, thereby establishing the groundwork for obtaining "artificial fish" in the future.
诱导多能干细胞（iPSCs）是一种新型的多能细胞，它由体细胞重新编程为胚胎样多能干细胞，用于研究发育、疾病和潜在的基因治疗。鱼类 iPSCs 的诱导和调控机制尚不清楚。我们通过转染技术，研究了 OSKMNL 因子共表达对大黄鱼（Larimichthys crocea）肌肉细胞系（LYCMs）体细胞重编程的关键功能，并成功建立了一个稳定的 iPSCs 系（Lc-OSNL-iPSCs）。实现了碱性磷酸酶活性高、染色体核型稳定的 iPSCs 稳定培养。qRT-PCR 和免疫荧光标记结果显示，Lc-OSNL-iPSCs 高表达多能标记基因如 Nanog、Oct4 和 Sox2。Lc-OSNL-iPSCs、Lc-OSKMNL-iPSCs 和 LYCMs 之间存在显著差异，当激活干细胞多能性信号通路时，维持细胞多能性的几个基因的表达上调。本研究构建了大黄鱼 iPSCs 的诱导技术体系。该系统可作为研究大黄鱼生殖细胞分化机制、性别控制、遗传和育种的基础模型，也可作为研究鱼类 iPSCs 的平台。有趣的是，获得的 iPSCs 可作为定向诱导肌肉干细胞的有用材料，为未来获得“人工鱼”奠定基础。
