Isolation and characterisation of exosomes from Chinese hamster ovary (CHO) cells.
Isolation and characterisation of exosomes from Chinese hamster ovary (CHO) cells.
复制标题
DOI：
10.1007/s10529-023-03353-3
复制
发表时间：
2023-04
影响因子：
2.7
通讯作者：
Smales, C. Mark
中科院分区：
文献类型：
--
作者：
Skrika-Alexopoulos, Eleftheria;Smales, C. Mark
外来体先前已经从中国仓鼠卵巢（CHO）细胞中分离，并且报道了它们的抗凋亡特性。然而，为了进一步促进CHO细胞衍生的外泌体的研究并允许它们在研究中进行比较，有必要使用至少三个标准来鉴定和定义这样的外泌体，所述标准可以用作CHO细胞产生的外泌体的产生的参考。在这里，我们报告了使用总外泌体分离（TEI）方法在分批培养期间生长的指数期和稳定期期间从CHO细胞中分离外泌体，CHO细胞是一种工业上相关且广泛使用的用于生物制药蛋白质生产的细胞宿主。使用各种技术，包括动态光散射（DLS），Zeta电位，电子显微镜和免疫印迹，并确定其蛋白质和RNA含量的特点和可视化所得的囊泡。我们还使用MALDI-ToF质谱法生成了分离的外泌体的脂质指纹。我们证实了来自CHO细胞的纳米尺寸的细胞外囊泡的存在，并且其随后的表征揭示了其大小、均匀性、表面电荷、蛋白质和RNA含量的细节。还发现外泌体的脂质含量在分批培养的不同天数分离的外泌体之间不同。该分析提供了CHO细胞外泌体的概况和表征，以帮助未来对来自CHO细胞的外泌体的研究，并改善用于生物制药应用的外泌体的制造。
Exosomes have previously been isolated from Chinese hamster ovary (CHO) cells and their anti-apoptotic properties reported. However, to further facilitate the study of CHO cell derived exosomes and allow their comparison across studies, it is necessary to characterise and define such exosomes using at least three criteria that can act as a reference for the generation of CHO cell produced exosomes. Here we report on the isolation of exosomes from CHO cells, an industrially relevant and widely used cell host for biopharmaceutical protein production, during the exponential and stationary phase of growth during batch culture using a Total Exosome Isolation (TEI) method. The resulting vesicles were characterized and visualized using a diverse range of techniques including Dynamic Light Scattering (DLS), Zeta potential, Electron Microscopy and immunoblotting, and their protein and RNA content determined. We also generated the lipid fingerprint of isolated exosomes using MALDI-ToF mass spectroscopy. We confirmed the presence of nano sized extracellular vesicles from CHO cells and their subsequent characterization revealed details of their size, homogeneity, surface charge, protein and RNA content. The lipid content of exosomes was also found to differ between exosomes isolated on different days of batch culture. This analysis provides a profile and characterisation of CHO cell exosomes to aid future studies on exosomes from CHO cells and improving the manufacturing of exosomes for biotherapeutic application.
登录
查看更多内容
影响因子：
16
作者：
Lötvall J;Hill AF;Hochberg F;Buzás EI;Di Vizio D;Gardiner C;Gho YS;Kurochkin IV;Mathivanan S;Quesenberry P;Sahoo S;Tahara H;Wauben MH;Witwer KW;Théry C
通讯作者：
Théry C
DOI：
10.1208/s12248-018-0227-4
发表时间：
2018-04-30
期刊：
The AAPS journal
影响因子：
--
作者：
Ferguson S;Kim S;Lee C;Deci M;Nguyen J
通讯作者：
Nguyen J
影响因子：
3.8
作者：
Han, Seora;Rhee, Won Jong
通讯作者：
Rhee, Won Jong
影响因子：
4.1
作者：
Povey, Jane F.;O'Malley, Christopher J.;Smales, C. Mark
通讯作者：
Smales, C. Mark
基于聚合物的沉淀保留了子宫内膜细胞系细胞外囊泡的生物活性。
影响因子：
3.7
作者：
Niu Z;Pang RTK;Liu W;Li Q;Cheng R;Yeung WSB
通讯作者：
Yeung WSB