Tissue engineering and cell transplantation therapy have become promising therapies for intractable diseases. These approaches require cell separation technology without cell modification. Accordingly, in this study, we developed a novel cell separation method using a thermoresponsive block copolymer brush with an affinity peptide. A block copolymer brush with bottom poly(2-hydroxyethyl methacrylate [HEMA]-co-propargyl acrylate) and top poly(N-isopropylacrylamide-co-HEMA) segments was prepared through two steps of atom transfer radical polymerization. Then, cell affinity peptides were conjugated to the bottom segment of the copolymer brush through a click reaction. Using cRGD as a cell-affinity peptide, enhancement of cell adhesion with rapid adhesion on the copolymer brush was observed at 37 °C, whereas the copolymer brush without cRGD did not exhibit cell adhesion. Temperature-modulated cell adhesion and detachment were performed with a relatively long upper segment because the affinity between peptides and cells was modulated by the swelling and shrinking of the upper thermoresponsive segment. Selective endothelial cell adhesion was performed at 37 °C using GGGREDV as an affinity peptide. Smooth muscle cells and fibroblasts did not adhere to the copolymer brush. Adhered human umbilical vein endothelial cells (HUVECs) were successfully recovered by reducing the temperature to 20 °C. Based on the properties of the copolymer brush, HUVECs could be purified using a mixture of cells simply by changing the temperature. These results demonstrated that the prepared copolymer brush with cell affinity peptides could be a useful cell separation tool because the cells could be separated with specificity and without cell modification using a simple procedure.

中文翻译：

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使用具有亲和力肽的热敏聚合物刷选择性捕获和非侵入性释放细胞

组织工程和细胞移植疗法已成为治疗顽固性疾病的有前途的疗法。这些方法需要细胞分离技术而无需细胞修饰。因此，在这项研究中，我们开发了一种使用具有亲和力肽的热响应性嵌段共聚物刷的新型细胞分离方法。的嵌段共聚物刷底部聚（2-羟乙基甲基丙烯酸酯[HEMA] -共-propargyl丙烯酸酯）和顶部聚（Ñ -isopropylacrylamide-共-HEMA）链段是通过两步原子转移自由基聚合制备的。然后，通过点击反应将细胞亲和肽缀合到共聚物刷的底部。使用cRGD作为细胞亲和肽，在37°C时观察到了在共聚物刷上的快速附着，细胞附着力得到了增强，而没有cRGD的共聚物刷则没有细胞附着。温度调节的细胞粘附和分离是用相对较长的上段进行的，因为肽和细胞之间的亲和力是通过上热响应段的膨胀和收缩来调节的。使用GGGREDV作为亲和力肽在37°C下进行选择性内皮细胞粘附。平滑肌细胞和成纤维细胞没有粘附到共聚物刷上。通过将温度降低至20°C，成功恢复了粘附的人脐静脉内皮细胞（HUVEC）。根据共聚物刷的特性，可以通过简单地改变温度，使用混合细胞来纯化HUVEC。这些结果表明，制备的具有细胞亲和力肽的共聚物刷可以是有用的细胞分离工具，因为可以使用简单的程序特异性地分离细胞而无需进行细胞修饰。