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从中国仓鼠肺组织中建立快速生长的无血清永生化细胞,用于生物制药生产。

Establishment of fast-growing serum-free immortalised cells from Chinese hamster lung tissues for biopharmaceutical production.

机构信息

Graduate School of Engineering, Osaka University, 2-1, Yamadaoka, Suita, Osaka, 565-0871, Japan.

Manufacturing Technology Association of Biologics, 7-1-49, Minatojima-Minamimachi, Chuo-ku, Kobe, Hyogo, 650-0047, Japan.

出版信息

Sci Rep. 2020 Oct 19;10(1):17612. doi: 10.1038/s41598-020-74735-0.

Abstract

Chinese hamster (Cricetulus griseus) ovary-derived Chinese hamster ovary (CHO) cells are the most commonly used mammalian hosts for the industrial production of recombinant therapeutics because of their ability to fold, assemble, and perform post-translational modifications, such as glycosylation, on proteins. They are also valuable for their ability to grow in serum-free suspension cultures. In this study, we established a cell line derived from lung tissue of Chinese hamsters, named Chinese hamster lung (CHL)-YN cells. The biosafety of CHL-YN cells was confirmed by in vitro sterility testing, mycoplasma detection, and reverse transcriptase assays. One of the key characteristics of CHL-YN cells was their doubling time of 8.1 h in chemically defined culture medium; thus, they proliferate much faster than conventional CHO cells and general mammalian cells. Transgenes could be introduced into CHL-YN cells with high efficiency. Finally, between 50% to > 100% of the amount of glycosylated immunoglobulin G (IgG)1 produced by CHO-K1 cells was produced by CHL-YN cells over a shorter period of time. In summary, fast-growing CHL-YN cells are a unique cell line for producing recombinant proteins.

摘要

中国仓鼠(Cricetulus griseus)卵巢来源的中国仓鼠卵巢(CHO)细胞因其能够折叠、组装和进行翻译后修饰,如糖基化等,成为工业生产重组治疗药物的最常用哺乳动物宿主。它们还因其能够在无血清悬浮培养中生长而具有重要价值。在这项研究中,我们建立了一个源自中国仓鼠肺组织的细胞系,命名为中国仓鼠肺(CHL)-YN 细胞。通过体外无菌试验、支原体检测和逆转录酶试验证实了 CHL-YN 细胞的生物安全性。CHL-YN 细胞的一个关键特征是在化学成分确定的培养基中的倍增时间为 8.1 小时;因此,它们比传统的 CHO 细胞和一般的哺乳动物细胞增殖得更快。转染基因可以高效地导入 CHL-YN 细胞。最后,在较短的时间内,CHL-YN 细胞产生的糖基化免疫球蛋白 G(IgG)1 的量是 CHO-K1 细胞的 50%至>100%。总之,生长迅速的 CHL-YN 细胞是生产重组蛋白的独特细胞系。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e322/7572389/0a808ba9ab1b/41598_2020_74735_Fig1_HTML.jpg

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