A single-cell surgery microfluidic device for transplanting tumor cytoplasm into dendritic cells without nuclei mixing
A single-cell surgery microfluidic device for transplanting tumor cytoplasm into dendritic cells without nuclei mixing
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DOI：
10.1002/biot.202200135
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发表时间：
2022-11-29
影响因子：
4.7
通讯作者：
Oana，Hidehiro
中科院分区：
文献类型：
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作者：
Okeyo，Kennedy Omondi;Hiyaji，Ryuta;Oana，Hidehiro
本研究旨在证明在集成一对一电融合、剪切流再分离和装置上培养的微流体装置中通过单细胞手术生成肿瘤细胞疫苗模型的可行性。该器械由聚二甲基硅氧烷（PDMS）制成，由用于一对一融合的微孔（孔径：1.3 μm）和用于器械上培养的微笼组成。使用该装置，我们可以实现白血病浆细胞样树突状细胞（DC样细胞）和Jurkat细胞的一对一电融合，融合效率为约80%。通过狭窄微孔的融合允许DC样细胞获得Jurkat细胞的细胞质内容物，同时防止核混合。融合后，通过剪切流应用将DC样细胞与Jurkat细胞选择性地再分离，以产生无肿瘤核的抗原受体DC样（tarDC样）细胞。当在装置上以单细胞形式培养时，这些细胞可以在温和的培养基灌注下存活，中位存活时间为11.5h，尽管少数细胞可以存活超过36h。总的来说，这项研究证明了微流体装置中的单细胞手术可能产生未被肿瘤核酸材料污染的树突状细胞疫苗。我们相信，这项研究将激发更安全的肿瘤细胞疫苗的产生，用于癌症免疫治疗。
This study aimed to demonstrate the feasibility of generating tumor cell vaccine models by single‐cell surgery in a microfluidic device that integrates one‐to‐one electrofusion, shear flow reseparation, and on‐device culture. The device was microfabricated from polydimethylsiloxane (PDMS) and consisted of microorifices (aperture size: ∼3 μm) for one‐to‐one fusion, and microcages for on‐device culture. Using the device, we could achieve one‐to‐one electrofusion of leukemic plasmacytoid dendritic cells (DC‐like cells) and Jurkat cells with a fusion efficiency of ∼ 80%. Fusion via the narrow microorifices allowed DC‐like cells to acquire cytoplasmic contents of the Jurkat cells while preventing nuclei mixing. After fusion, the DC‐like cells were selectively reseparated from the Jurkat cells by shear flow application to generate tumor nuclei‐free antigen‐recipient DC‐like (tarDC‐like) cells. When cultured as single cells on the device, these cells could survive under gentle medium perfusion with a median survival time of 11.5 h, although a few cells could survive longer than 36 h. Overall, this study demonstrates single‐cell surgery in a microfluidic device for potential generation of dendritic cell vaccines which are uncontaminated with tumor nucleic materials. We believe that this study will inspire the generation of safer tumor cell vaccines for cancer immunotherapy.