Inhibition of apoptosis using exosomes in Chinese hamster ovary cell culture
Inhibition of apoptosis using exosomes in Chinese hamster ovary cell culture
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DOI：
10.1002/bit.26549
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发表时间：
2018-05-01
影响因子：
3.8
通讯作者：
Rhee, Won Jong
中科院分区：
文献类型：
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作者：
Han, Seora;Rhee, Won Jong
在过去的几十年里，用于治疗性蛋白生产的动物细胞培养技术有了显著的进步。中国仓鼠卵巢（CHO）细胞已广泛应用于生物制药生产。在生物制药工业中，开发细胞培养基和培养条件以达到最高的生产力和质量是至关重要的。然而，CHO细胞在生物反应器中受到凋亡的显著影响，导致产品数量和质量大幅下降。因此，为了克服CHO细胞培养中凋亡的障碍，开发一种不考虑安全性和成本的新方法至关重要。在此，我们首次证明了外泌体（一种纳米级的细胞外囊泡）来源于CHO细胞，当添加到培养基中时，可以抑制CHO细胞培养中的凋亡。流式细胞术和显微镜分析显示，大量外泌体被递送到CHO细胞。与对照组（41.1%）相比，补充外泌体（67.3%）的staurosporine处理后的细胞存活率更高。此外，外泌体阻止线粒体膜电位丢失和caspase-3激活，这意味着外泌体增强了促凋亡条件下的细胞活性。由于外泌体补充剂来源于CHO细胞本身，因此它不仅有利于CHO细胞培养抑制凋亡的生物制药生产力，而且从监管的角度来看，也可以减少任何安全问题。因此，我们得出结论，本研究中开发的方法可能有助于生物制药工业，其中最大限度地减少CHO细胞培养中的凋亡是有益的。
Animal cell culture technology for therapeutic protein production has shown significant improvement over the last few decades. Chinese hamster ovary (CHO) cells have been widely adapted for the production of biopharmaceutical drugs. In the biopharmaceutical industry, it is crucial to develop cell culture media and culturing conditions to achieve the highest productivity and quality. However, CHO cells are significantly affected by apoptosis in the bioreactors, resulting in a substantial decrease in product quantity and quality. Thus, to overcome the obstacle of apoptosis in CHO cell culture, it is critical to develop a novel method that does not have minimal concern of safety or cost. Herein, we showed for the first time that exosomes, which are nano-sized extracellular vesicles, derived from CHO cells inhibited apoptosis in CHO cell culture when supplemented to the culture medium. Flow cytometric and microscopic analyses revealed that substantial amounts of exosomes were delivered to CHO cells. Higher cell viability after staurosporine treatment was observed by exosome supplementation (67.3%) as compared to control (41.1%). Furthermore, exosomes prevented the mitochondrial membrane potential loss and caspase-3 activation, meaning that the exosomes enhanced cellular activities under pro-apoptotic condition. As the exosomes supplements are derived from CHO cells themselves, it is not only beneficial for the biopharmaceutical productivity of CHO cell culture to inhibit apoptosis, but also from a regulatory standpoint to diminish any safety concerns. Thus, we conclude that the method developed in this research may contribute to the biopharmaceutical industry where minimizing apoptosis in CHO cell culture is beneficial.