Surface design of antibody-immobilized thermoresponsive cell culture dishes for recovering intact cells by low-temperature treatment
Surface design of antibody-immobilized thermoresponsive cell culture dishes for recovering intact cells by low-temperature treatment
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用于通过低温处理回收完整细胞的抗体固定化热敏细胞培养皿的表面设计
DOI：
10.1002/jbm.a.35064
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通讯作者：
Okano T
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作者：
Kobayashi J;Hayashi M;Ohno T;Nishi M;Arisaka Y;Matsubara Y;Kakidachi H;Akiyama Y;Yamato M;Horii A;Okano T
抗体固定化的温敏性聚（N-异丙基丙烯酰胺-co-2-羧基异丙基丙烯酰胺）[聚（IPAAm-co-CIPAAm）]接枝细胞培养表面被设计为通过调节细胞表面上固定化抗体和抗原之间的亲和力结合来增强弱粘附细胞的初始粘附和细胞响应低温而分离的能力。在细胞表面表达CD 90的Ty-82细胞和新生儿正常人真皮成纤维细胞（NHDF）在37°C下粘附于固定有抗CD 90抗体的温敏表面，在此条件下，移植的温敏聚合物链收缩。通过将温度降低至20°C并施加外力（例如移液），使贴壁Ty-82细胞从表面脱离，而培养的NHDF片仅响应于降低的温度而自发地从表面脱离。当温度降至20°C时，由于固定化抗体的抗原识别位点周围的聚合物链的空间位阻增加，接枝的温敏性聚合物链的溶胀减弱了固定化抗体和细胞上抗原之间的亲和力结合。通过低温处理从培养表面上共价固定的抗体收获的细胞上未检测到污染，而观察到抗体和抗生物素蛋白从抗生物素蛋白-生物素结合表面的残留。此外，在抗体固定的温度响应表面上，脂肪组织来源的细胞的初始粘附（其与PIPAAm接枝表面的粘附较弱）得到增强。© 2013 Wiley Periodicals，Inc. J Biomed Mater Res Part A：102A：3883-3893，2014.
Antibody‐immobilized thermoresponsive poly(N‐isopropylacrylamide‐co‐2‐carboxyisopropylacrylamide) [poly(IPAAm‐co‐CIPAAm)]‐grafted cell culture surfaces were designed to enhance both the initial adhesion of weakly adhering cells and the ability of cells to detach in response to low temperature through the regulation of affinity binding between immobilized antibodies and antigens on the cellular surface. Ty‐82 cells and neonatal normal human dermal fibroblasts (NHDFs), which express CD90 on the cell surface, adhered to anti‐CD90 antibody‐immobilized thermoresponsive surfaces at 37°C, a condition at which the grafted thermoresponsive polymer chains shrank. Adherent Ty‐82 cells were detached from the surfaces by lowering the temperature to 20°C and applying external forces, such as pipetting, whereas cultured NHDF sheets spontaneously detached themselves from the surface in response to reduced temperature alone. When the temperature was decreased to 20°C, the swelling of grafted thermoresponsive polymer chains weakened the affinity binding between immobilized antibody and antigen on the cells due to the increasing steric hindrance of the polymer chains around the antigen‐recognition site of the immobilized antibodies. No contamination was detected on cells harvested from covalently immobilized antibodies on the culture surfaces by low‐temperature treatment, whereas a carryover of the antibody and avidin from the avidin‐biotin binding surface was observed. Furthermore, the initial adhesion of adipose tissue‐derived cells, which adhere weakly to PIPAAm‐grafted surfaces, was enhanced on the antibody‐immobilized thermoresponsive surfaces. © 2013 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 102A: 3883–3893, 2014.