Investigation of scaffold manufacturing conditions for 3-dimensional culture of myogenic cell line derived from black sea bream (Acanthopagrus schlegelii).

IF 2 4区 生物学 Q3 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Cytotechnology Pub Date : 2025-02-01 Epub Date: 2024-12-12 DOI:10.1007/s10616-024-00676-5
Ye-Eun Lee, Eun Soo Jeong, Young-Mog Kim, Seung Pyo Gong
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Abstract

Culturing fish myogenic cells in vitro holds significant potential to revolutionize aquaculture practices and support sustainable food production. However, advancement in in vitro culture technologies for skeletal muscle-derived myogenic cells have predominantly focused on mammals, with limited studies on fish. Scaffold-based three-dimensional (3D) culture systems for fish myogenic cells remain underexplored, highlighting a critical research gap compared to mammalian systems. This study evaluated the effects of scaffold composition and manufacturing methods on cellular growth in the 3D culture of black sea bream (Acanthopagrus schlegelii) myogenic cells. Scaffolds were manufactured using three natural polymers: black sea bream-derived extracellular matrix (ECM), sodium alginate, and gelatin. Two scaffold types were tested: "cell-laden scaffolds" prepared by mixing cells into the pre-scaffold solution followed by gelation, and "cell-seeding scaffolds" produced by freezing, gelation, and lyophilization before cell inoculation. Scaffold characteristics, including pore size, porosity, swelling ratio, and degradation rate, were assessed. Cell-seeding scaffolds exhibited relatively larger pore size, higher porosity, and higher degradation rate, while cell-laden scaffolds had higher swelling ratios. When black sea bream myogenic cells were cultured in these scaffolds, cell-seeding scaffolds supported cellular growth, particularly when composed of 3% sodium alginate and 4% gelatin with any concentration of ECM. In contrast, cell-laden scaffolds did not support cellular growth regardless of their composition. These findings provide fundamental insights for optimizing scaffold properties to develop more optimized conditions for 3D culture of fish muscle lineage cells.

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研究从黑鲷(Acanthopagrus schlegelii)中提取的肌原细胞系的三维培养支架制造条件。
体外培养鱼类肌原细胞在革新水产养殖方法和支持可持续食品生产方面具有巨大潜力。然而,骨骼肌肌原细胞体外培养技术的进展主要集中在哺乳动物上,对鱼类的研究十分有限。基于支架的鱼类肌原细胞三维(3D)培养系统仍未得到充分探索,这凸显了与哺乳动物系统相比存在的关键研究空白。本研究评估了黑鲷(Acanthopagrus schlegelii)肌原细胞三维培养中支架成分和制造方法对细胞生长的影响。支架由三种天然聚合物制成:源于黑鲷的细胞外基质(ECM)、海藻酸钠和明胶。测试了两种类型的支架:将细胞混合到预支架溶液中然后凝胶化制备的 "细胞负载支架",以及在细胞接种前通过冷冻、凝胶化和冻干制备的 "细胞播种支架"。评估了支架的特性,包括孔径、孔隙率、膨胀率和降解率。细胞接种支架的孔径相对较大,孔隙率较高,降解率较高,而细胞负载支架的膨胀率较高。在这些支架中培养黑鲷成肌细胞时,细胞播撒支架支持细胞生长,尤其是由 3% 的海藻酸钠和 4% 的明胶以及任何浓度的 ECM 组成的支架。相比之下,无论其成分如何,含有细胞的支架都不支持细胞生长。这些发现为优化支架特性提供了基本见解,从而为鱼类肌肉系细胞的三维培养提供更优化的条件。
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来源期刊
Cytotechnology
Cytotechnology 生物-生物工程与应用微生物
CiteScore
4.10
自引率
0.00%
发文量
49
审稿时长
6-12 weeks
期刊介绍: The scope of the Journal includes: 1. The derivation, genetic modification and characterization of cell lines, genetic and phenotypic regulation, control of cellular metabolism, cell physiology and biochemistry related to cell function, performance and expression of cell products. 2. Cell culture techniques, substrates, environmental requirements and optimization, cloning, hybridization and molecular biology, including genomic and proteomic tools. 3. Cell culture systems, processes, reactors, scale-up, and industrial production. Descriptions of the design or construction of equipment, media or quality control procedures, that are ancillary to cellular research. 4. The application of animal/human cells in research in the field of stem cell research including maintenance of stemness, differentiation, genetics, and senescence, cancer research, research in immunology, as well as applications in tissue engineering and gene therapy. 5. The use of cell cultures as a substrate for bioassays, biomedical applications and in particular as a replacement for animal models.
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