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利用喷射注射器施加瞬态压力提高聚乙二醇细胞融合效率

Enhancement of polyethylene glycol-cell fusion efficiency by novel application of transient pressure using a jet injector.

机构信息

Department of Device Application for Molecular Therapeutics, Graduate School of Medicine, Osaka University, Japan.

Medical Device Division, Industry Business Unit, Safety Strategic Business Unit, Daicel Co., Osaka, Japan.

出版信息

FEBS Open Bio. 2023 Mar;13(3):478-489. doi: 10.1002/2211-5463.13557. Epub 2023 Jan 27.

Abstract

Cell-cell fusion involves the fusion of somatic cells into a single hybrid cell. It is not only a physiological process but also an important cell engineering technology which can be applied to various fields, such as regenerative medicine, antibody engineering, genetic engineering, and cancer therapy. There are three major methods of cell fusion: electrical cell fusion, polyethylene glycol (PEG) cell fusion, and virus-mediated cell fusion. Although PEG cell fusion is the most economical approach and does not require expensive instrumentation, it has a poor fusion rate and induces a high rate of cell cytotoxicity. To improve the fusion rate of the PEG method, we combined it with the pyro-drive jet injector (PJI). PJI provides instant pressure instead of cell agitation to increase the probability of cell-to-cell contact and shorten the distance between cells in the process of cell fusion. Here, we report that this improved fusion method not only decreased cell cytotoxicity during the fusion process, but also increased fusion rate compared with the conventional PEG method. Furthermore, we tested the functionality of cells fused using the PJI-PEG method and found them to be comparable to those fused using the conventional PEG method in terms of their application for dendritic cell (DC)-tumor cell fusion vaccine production; in addition, the PJI-PEG method demonstrated excellent performance in hybridoma cell preparation. Taken together, our data indicate that this method improves cell fusion efficiency as compared to the PEG method and thus has the potential for use in various applications that require cell fusion technology.

摘要

细胞融合涉及体细胞融合成单个杂交细胞。它不仅是一个生理过程,也是一种重要的细胞工程技术,可应用于再生医学、抗体工程、基因工程和癌症治疗等各个领域。细胞融合有三种主要方法:电融合、聚乙二醇(PEG)融合和病毒介导的细胞融合。虽然 PEG 融合是最经济的方法,不需要昂贵的仪器,但融合率低,细胞毒性高。为了提高 PEG 法的融合率,我们将其与热驱动射流注射器(PJI)相结合。PJI 提供瞬间压力而不是细胞搅拌,以增加细胞间接触的概率,并在细胞融合过程中缩短细胞间的距离。在这里,我们报告说,这种改进的融合方法不仅降低了融合过程中的细胞毒性,而且与传统的 PEG 方法相比,融合率也有所提高。此外,我们测试了使用 PJI-PEG 方法融合的细胞的功能,发现它们在用于树突状细胞(DC)-肿瘤细胞融合疫苗生产方面与使用传统 PEG 方法融合的细胞相当;此外,PJI-PEG 方法在杂交瘤细胞制备方面表现出优异的性能。总之,我们的数据表明,与 PEG 法相比,该方法提高了细胞融合效率,因此有可能应用于需要细胞融合技术的各种应用。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5b86/9989930/74e3ea3c0e66/FEB4-13-478-g003.jpg

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