Induction of Pluripotent Stem Cells from Muscle Cells of Large Yellow Croaker (Larimichthys Crocea) Via Electrotransfection

IF 2.6 3区生物学 Q3 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Marine Biotechnology Pub Date : 2024-09-09 DOI:10.1007/s10126-024-10372-2

Zhaowei Zhong, Yilei Wang, Yan Feng, Yan Xu, Pengfei Zou, Ziping Zhang, Yonghua Jiang

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Abstract

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.

Graphical Abstract

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通过电转染从大黄鱼（Larimichthys Crocea）肌肉细胞中诱导多能干细胞。

诱导多能干细胞（iPSCs）是一种新型多能细胞，可将体细胞重编程为类似胚胎的多能干细胞状态，用于研究发育、疾病和潜在的基因疗法。鱼类 iPSCs 的诱导和调控机制尚不清楚。通过转染技术，我们研究了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作用，并成功建立了稳定的iPSCs细胞系（Lc-OSNL-iPSCs）。培养出的 iPSCs 具有较高的碱性磷酸酶活性和稳定的核型。qRT-PCR 和免疫荧光标记结果显示，Lc-OSNL-iPSCs 的多能标记基因（如 Nanog、Oct4 和 Sox2）表达水平较高。Lc-OSNL-iPSCs、Lc-OSKMNL-iPSCs和LYCMs之间存在明显差异，当干细胞多能性信号通路被激活时，维持细胞多能性的几个基因的表达被上调。本研究构建了诱导大菱鲆 iPSCs 的技术体系。该系统可作为了解大黄鱼生殖细胞分化机制、性别控制、遗传学和育种的基础模型，以及研究鱼类 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，从而为将来获得 "人造鱼 "奠定基础。

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来源期刊

Marine Biotechnology 工程技术-海洋与淡水生物学

CiteScore

4.80

自引率

3.30%

发文量

审稿时长

2 months

期刊介绍： Marine Biotechnology welcomes high-quality research papers presenting novel data on the biotechnology of aquatic organisms. The journal publishes high quality papers in the areas of molecular biology, genomics, proteomics, cell biology, and biochemistry, and particularly encourages submissions of papers related to genome biology such as linkage mapping, large-scale gene discoveries, QTL analysis, physical mapping, and comparative and functional genome analysis. Papers on technological development and marine natural products should demonstrate innovation and novel applications.