Automated adherent cell elimination by a high-speed laser mediated by a light-responsive polymer
Automated adherent cell elimination by a high-speed laser mediated by a light-responsive polymer
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DOI：
10.1038/s42003-018-0222-4
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发表时间：
2018-01-01
影响因子：
5.9
通讯作者：
Sumaru, Kimio
中科院分区：
文献类型：
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作者：
Hayashi, Yohei;Matsumoto, Junichi;Sumaru, Kimio
传统的细胞处理和分选方法需要手工劳动，这降低了细胞的质量和数量。为了纯化贴壁培养的细胞，期望高通量而不解离并且可以按需方式利用的细胞纯化技术。在这里，我们开发了一种激光诱导的光响应聚合物激活的细胞杀伤（LiLACK）系统，该系统能够实现高速和按需贴壁细胞切片和纯化。该系统采用可见激光束，其不直接杀死细胞，而是通过偶氮苯部分的反-顺-反光异构化诱导局部产热。在每次传代中使用该系统进行切片，人诱导多能干细胞（hiPSC）在长期培养期间保持其多能性和自我更新。此外，结合对荧光和相衬图像的深度机器学习分析，通过在未分化的hiPSC培养条件下消除不需要的自发分化细胞，开发了一种无标记的自动细胞处理系统。
Conventional cell handling and sorting methods require manual labor, which decreases both cell quality and quantity. To purify adherent cultured cells, cell purification technologies that are high throughput without dissociation and can be utilized in an on-demand manner are expected. Here, we developed a Laser-induced, Light-responsive-polymer-Activated, Cell Killing (LiLACK) system that enables high-speed and on-demand adherent cell sectioning and purification. This system employs a visible laser beam, which does not kill cells directly, but induces local heat production through the trans-cis-trans photo-isomerization of azobenzene moieties. Using this system in each passage for sectioning, human induced pluripotent stem cells (hiPSCs) maintained their pluripotency and self-renewal during long-term culture. Furthermore, combined with deep machine-learning analysis on fluorescent and phase contrast images, a label-free and automatic cell processing system has been developed by eliminating unwanted spontaneously differentiated cells in undifferentiated hiPSC culture conditions.