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用于温度调节干细胞分离的热响应性阳离子嵌段共聚物刷

Thermoresponsive Cationic Block Copolymer Brushes for Temperature-Modulated Stem Cell Separation.

作者信息

Nagase Kenichi, Ota Ayumu, Hirotani Tadashi, Yamada Sota, Akimoto Aya Mizutani, Kanazawa Hideko

机构信息

Faculty of Pharmacy, Keio University, 1-5-30 Shibakoen, Minato, Tokyo, 105-8512, Japan.

Department of Materials Engineering, Schools of Engineering, The University of Tokyo, 7-3-1 Hongo, Bunkyo, Tokyo, 113-8656, Japan.

出版信息

Macromol Rapid Commun. 2020 Oct;41(19):e2000308. doi: 10.1002/marc.202000308. Epub 2020 Aug 17.

Abstract

Recently, cell separation methods have become important for preparing cells for transplantation therapy. In this study, a thermoresponsive cationic block copolymer brush is developed as an effective cell separation tool. This brush is prepared on glass surfaces using two steps of activator regenerated by electron transfer-atom transfer radical polymerization (ARGET-ATRP). The cationic segment is prepared in the first step of the ARGET-ATRP of N,N-dimethylaminopropylacrylamide (DMAPAAm). In the second step, the thermoresponsive segment is prepared, attached to the bottom cationic segment, through ARGET-ATRP with N-isopropylacrylamide (NIPAAm). The cell adhesion behavior of the prepared thermoresponsive cationic copolymer, PDMAPAAm-b-PNIPAAm, brush is observed using umbilical cord-derived mesenchymal stem cells (UCMSC), fibroblasts, and macrophages. At 37 °C, all three types of cells adhere to the thermoresponsive cationic copolymer brush. Then, by reducing the temperature to 20 °C, the adhered UCMSC are detached from the copolymer brush, whereas the fibroblasts and macrophages remain adhered to the copolymer brush. Using this copolymer brush, UCMSC can be purified from the cell mixture simply by changing the temperature. Therefore, the prepared thermoresponsive cationic copolymer brush is useful as a cell separation tool for the purification of mesenchymal stem cells.

摘要

近年来,细胞分离方法对于为移植治疗制备细胞变得至关重要。在本研究中,开发了一种热响应性阳离子嵌段共聚物刷作为一种有效的细胞分离工具。该刷通过电子转移-原子转移自由基聚合(ARGET-ATRP)再生的活化剂两步法在玻璃表面制备。阳离子段在N,N-二甲基氨基丙基丙烯酰胺(DMAPAAm)的ARGET-ATRP第一步中制备。在第二步中,通过与N-异丙基丙烯酰胺(NIPAAm)的ARGET-ATRP制备热响应段,并将其连接到底部阳离子段上。使用脐带间充质干细胞(UCMSC)、成纤维细胞和巨噬细胞观察制备的热响应性阳离子共聚物PDMAPAAm-b-PNIPAAm刷的细胞粘附行为。在37℃时,所有三种类型的细胞都粘附在热响应性阳离子共聚物刷上。然后,通过将温度降低到20℃,粘附的UCMSC从共聚物刷上脱离,而成纤维细胞和巨噬细胞仍然粘附在共聚物刷上。使用这种共聚物刷,只需改变温度就可以从细胞混合物中纯化UCMSC。因此,制备的热响应性阳离子共聚物刷可作为一种用于纯化间充质干细胞的细胞分离工具。

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