Enhancement of intracellular extraction from Oscillatoria okine and the potential use of the extract as a supplement to fetal bovine serum in animal cell culture

IF 4.6 2区 生物学 Q1 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Algal Research-Biomass Biofuels and Bioproducts Pub Date : 2024-09-15 DOI:10.1016/j.algal.2024.103713
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Abstract

A robust cell wall and well-organized thylakoid cyanobacteria can be candidates as a promising resource for C-phycocyanin (C-PC) and intracellular applications in various biotechnological areas. However, the development of extraction techniques with minimal chemical contamination to obtain the components remains an ongoing challenge. This study aimed to assess the efficacy of C-PC and intracellular extraction from Ocillatoria okeni TISTR8549 utilizing freezing-thaw (FT), pulsed electric field (PEF), and high-pressure homogenization (HPH) techniques. Additionally, the potential of O. okeni extract as a supplement and substitute for fetal bovine serum (FBS) in HaCaT cell culture was investigated. FT appeared to be the most proper method for C-PC extraction, yielding the highest purity and yield, although requiring 18 cycles for product accomplishment. PEF seemed unsuitable for intracellular component extraction from cyanobacteria with thick cell walls. The need for increased pulses resulted in thermal elevation and prolonged incubation times led to protein degradation. HPH proves to be an effective method for intracellular extraction, yielding high protein content suitable for the potential substitution of FBS in mammalian cell culture. Particularly, increasing pressure during HPH extraction leads to a decrease in protein yield. Resazurin and SRB assays revealed that adding algal extract in culture medium at concentrations of 1 % (w/v) improved HaCaT cell viability without disrupting cell morphology and metabolic processes. However, substituting algal extract in FBS resulted in cell proliferation decrease. Therefore, supplementing O. okeni extract during cell culture improved HaCaT cell proliferation, but it was unsuitable substitute for FBS in the culture medium. However, the feasibility of employing algal extracts as FBS replacements in cell culture is interesting and warrants further detailed investigation into the specific intracellular components that could serve as substitutes for FBS. Such an approach could offer an alternative source, mitigating ethical concerns and reducing costs associated with FBS usage.

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提高鹅膏蕈细胞内提取物的提取率以及将其用作动物细胞培养中胎牛血清补充剂的可能性
蓝藻具有坚固的细胞壁和良好的组织结构,可作为 C-植物蓝藻素(C-PC)和细胞内蓝藻素在各种生物技术领域应用的理想资源。然而,开发化学污染最小的提取技术以获得这些成分仍是一项持续的挑战。本研究旨在利用冷冻-解冻(FT)、脉冲电场(PEF)和高压匀浆(HPH)技术,评估从 Ocillatoria okeni TISTR8549 中提取 C-PC 和细胞内成分的功效。此外,还研究了 O. okeni 提取物在 HaCaT 细胞培养中作为胎牛血清(FBS)的补充和替代品的潜力。FT 似乎是提取 C-PC 的最合适方法,纯度和产量都最高,但需要 18 个循环才能获得产品。PEF 似乎不适合从细胞壁较厚的蓝藻中提取细胞内成分。需要增加脉冲导致热量升高,延长培养时间导致蛋白质降解。事实证明,HPH 是一种有效的细胞内提取方法,其蛋白质含量高,可用于替代哺乳动物细胞培养中的 FBS。特别是在 HPH 提取过程中,压力的增加会导致蛋白质产量的下降。利马唑啉和 SRB 检测显示,在培养基中添加浓度为 1%(重量/体积)的海藻提取物可提高 HaCaT 细胞的活力,同时不会破坏细胞形态和代谢过程。然而,用 FBS 代替海藻提取物会导致细胞增殖减少。因此,在细胞培养过程中补充海藻提取物能提高 HaCaT 细胞的增殖能力,但它并不适合替代培养基中的 FBS。然而,在细胞培养中使用海藻提取物作为 FBS 替代品的可行性非常有趣,值得进一步详细研究可作为 FBS 替代品的特定细胞内成分。这种方法可以提供一种替代来源,减轻道德方面的担忧,并降低与使用 FBS 相关的成本。
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来源期刊
Algal Research-Biomass Biofuels and Bioproducts
Algal Research-Biomass Biofuels and Bioproducts BIOTECHNOLOGY & APPLIED MICROBIOLOGY-
CiteScore
9.40
自引率
7.80%
发文量
332
期刊介绍: Algal Research is an international phycology journal covering all areas of emerging technologies in algae biology, biomass production, cultivation, harvesting, extraction, bioproducts, biorefinery, engineering, and econometrics. Algae is defined to include cyanobacteria, microalgae, and protists and symbionts of interest in biotechnology. The journal publishes original research and reviews for the following scope: algal biology, including but not exclusive to: phylogeny, biodiversity, molecular traits, metabolic regulation, and genetic engineering, algal cultivation, e.g. phototrophic systems, heterotrophic systems, and mixotrophic systems, algal harvesting and extraction systems, biotechnology to convert algal biomass and components into biofuels and bioproducts, e.g., nutraceuticals, pharmaceuticals, animal feed, plastics, etc. algal products and their economic assessment
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