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适应无血清培养条件的细胞系。

Adapting Cell Lines to Serum-Free Culture Conditions.

机构信息

Drosophila Genomics Resource Center, Indiana University, Bloomington, Indiana 47405.

Department of Biology, Indiana University, Bloomington, Indiana 47405.

出版信息

G3 (Bethesda). 2020 Dec 3;10(12):4541-4551. doi: 10.1534/g3.120.401769.

Abstract

Successful cell culture relies on media containing xenogenic components such as fetal bovine serum to support continuous cell proliferation. Here, we report a serum-free culture condition that supports the growth and proliferation of S2R+ and Kc167 cell lines. Importantly, the gradual adaptation of S2R+ and Kc167 cells to a media lacking serum was supported by supplementing the media with adult soluble extract, commonly known as fly extract. The utility of these adapted cells lines is largely unchanged. The adapted cells exhibited robust proliferative capacity and a transfection efficiency that was comparable to control cells cultured in serum-containing media. Transcriptomic data indicated that the S2R+ cells cultured with fly extract retain their hemocyte-specific transcriptome profile, and there were no global changes in the transcriptional output of cell signaling pathways. Our metabolome studies indicate that there were very limited metabolic changes. In fact, the cells were likely experiencing less oxidative stress when cultured in the serum-free media supplemented with fly extract. Overall, the cell culture conditions reported here consequently provide researchers with an alternative and physiologically relevant resource to address cell biological research questions.

摘要

成功的细胞培养依赖于含有异种成分(如胎牛血清)的培养基来支持细胞的连续增殖。在这里,我们报告了一种无血清培养条件,可支持 S2R+和 Kc167 细胞系的生长和增殖。重要的是,通过在培养基中添加成人可溶性提取物(通常称为果蝇提取物),逐步适应无血清培养基可以支持 S2R+和 Kc167 细胞的生长。这些适应细胞系的用途基本保持不变。适应细胞表现出强大的增殖能力和转染效率,与在含血清培养基中培养的对照细胞相当。转录组数据表明,用果蝇提取物培养的 S2R+细胞保留了其血细胞特异性转录组特征,细胞信号通路的转录输出没有全局变化。我们的代谢组学研究表明代谢变化非常有限。事实上,当在补充有果蝇提取物的无血清培养基中培养时,细胞可能经历的氧化应激更少。总的来说,这里报道的细胞培养条件为研究人员提供了一种替代的、生理相关的资源,以解决细胞生物学研究问题。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7f56/7718738/802bfb85b4a3/4541f1.jpg

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