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动物成分-free 细胞培养过渡案例研究。

Case Studies Exemplifying the Transition to Animal Component-free Cell Culture.

机构信息

84510Animal Welfare Academy of the German Animal Welfare Federation, Neubiberg, Germany.

608172Cellasys GmbH, Kronburg, Germany.

出版信息

Altern Lab Anim. 2022 Sep;50(5):330-338. doi: 10.1177/02611929221117999. Epub 2022 Aug 19.

Abstract

Cell culture techniques are strongly connected with modern scientific laboratories and production facilities. Thus, choosing the most suitable medium for the cells involved is vital, not only directly to optimise cell viability but also indirectly to maximise the reliability of the experiments performed with the cells. Fetal bovine or calf serum (FBS or FCS, respectively) is the most commonly used cell culture medium supplement, providing various nutritional factors and macromolecules essential for cell growth. Yet, the use of FBS encompasses a number of disadvantages. Scientifically, one of the most severe disadvantages is the lot-to-lot variability of animal sera that hampers reproducibility. Therefore, transitioning from the use of these ill-defined, component-variable, inconsistent, xenogenic, ethically questionable and even potentially infectious media supplements, is key to achieving better data reproducibility and thus better science. To demonstrate that the transition to animal component-free cell culture is possible and achievable, we highlight three different scenarios and provide some case studies of each, namely: i) the adaptation of single cell lines to animal component-free culture conditions by the replacement of FBS and trypsin; ii) the adaptation of multicellular models to FBS-free conditions; and (iii) the replacement of FBS with human platelet lysate (hPL) for the generation of primary stem/stromal cell cultures for clinical purposes. By highlighting these examples, we aim to foster and support the global movement towards more consistent science and provide evidence that it is indeed possible to step out of the currently smouldering scientific reproducibility crisis.

摘要

细胞培养技术与现代科学实验室和生产设施密切相关。因此,选择最适合所涉及细胞的培养基至关重要,不仅直接优化细胞活力,而且间接最大化用细胞进行的实验的可靠性。胎牛或小牛血清(分别为 FBS 或 FCS)是最常用的细胞培养基补充物,提供各种营养因子和大分子,是细胞生长所必需的。然而,FBS 的使用存在许多缺点。从科学角度来看,最严重的缺点之一是动物血清的批次间变异性,这阻碍了可重复性。因此,从使用这些定义不明确、成分可变、不一致、异源、存在伦理问题甚至潜在传染性的培养基补充物过渡,是实现更好的数据重现性从而获得更好的科学的关键。为了证明向无动物成分的细胞培养过渡是可能和可行的,我们突出了三种不同的情况,并提供了每种情况的一些案例研究,即:i)通过替代 FBS 和胰蛋白酶来适应单细胞系的无动物成分培养条件;ii)适应多细胞模型的无 FBS 条件;和 iii)用人血小板裂解物(hPL)替代 FBS 用于生成用于临床目的的原代干细胞/基质细胞培养物。通过突出这些例子,我们旨在促进和支持全球朝着更一致的科学发展,并提供证据表明,确实有可能走出当前弥漫着的科学可重复性危机。

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