PDF(Pubmed)
Abstract：
Cell culturing methods are increasingly used to reduce and replace the use of live animals in biomedical research and chemical toxicity testing. Although live animals are avoided when using cell culturing methods, they often contain animal-derived components of which one of the most commonly used is foetal bovine serum (FBS). FBS is added to cell culture media among other supplements to support cell attachment/spreading and cell proliferation. The safety, batch-to-batch variation, and ethical problems with FBS are acknowledged and therefore world-wide efforts are ongoing to produce FBS free media. Here, we present the composition of a new defined medium with only human proteins either recombinant or derived from human tissues. This defined medium supports long-term culturing/routine culturing of normal cells and of cancer cells, and can be used for freezing and thawing of cells, i.e. for cell banking. Here, we show for our defined medium, growth curves and dose response curves of cells grown in two and three dimensions, and applications such as cell migration. Cell morphology was studied in real time by phase contrast and phase holographic microscopy time-lapse imaging. The cell lines used are human cancer-associated fibroblasts, keratinocytes, breast cancer JIMT-1 and MDA-MB-231 cells, colon cancer CaCo-2 cells, and pancreatic cancer MiaPaCa-2 cells as well as the mouse L929 cell line. In conclusion, we present the composition of a defined medium without animal-derived products which can be used for routine culturing and in experimental settings for normal cells and for cancer cells, i.e. our defined medium provides a leap towards a universal animal product free cell culture medium.
摘要：
细胞培养方法越来越多地用于减少和替代活体动物在生物医学研究和化学毒性测试中的使用。尽管在使用细胞培养方法时避免了活体动物，它们通常含有动物来源的成分，其中最常用的是胎牛血清（FBS）。将FBS添加至细胞培养基以及其他补充物以支持细胞附着/扩散和细胞增殖。安全,批次到批次的变化，FBS的道德问题得到了承认，因此全世界都在努力生产FBS免费媒体。这里,我们提供了一种新的定义培养基的组合物，该培养基仅具有重组或源自人体组织的人类蛋白质。这种确定的培养基支持正常细胞和癌细胞的长期培养/常规培养，可用于细胞的冷冻和解冻，即用于细胞库。这里,我们展示了我们定义的媒介，在二维和三维生长的细胞的生长曲线和剂量反应曲线，以及细胞迁移等应用。通过相衬和相位全息显微镜延时成像实时研究细胞形态。使用的细胞系是人类癌症相关的成纤维细胞，角质形成细胞，乳腺癌JIMT-1和MDA-MB-231细胞，结肠癌CaCo-2细胞，和胰腺癌MiaPaCa-2细胞以及小鼠L929细胞系。总之,我们提供了一种不含动物衍生产品的确定培养基的组合物，可用于常规培养和正常细胞和癌细胞的实验设置，即我们的确定培养基提供了向通用动物产品无细胞培养基的飞跃。
