The use of patterned dual thermoresponsive surfaces for the collective recovery as co-cultured cell sheets
The use of patterned dual thermoresponsive surfaces for the collective recovery as co-cultured cell sheets
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DOI：
10.1016/j.biomaterials.2004.06.005
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发表时间：
2005-05-01
期刊：
影响因子：
14
通讯作者：
Okano, T
中科院分区：
文献类型：
--
作者：
Tsuda, Y;Kikuchi, A;Okano, T
异型细胞相互作用对于在许多组织和器官中实现和维持特定功能至关重要。我们专注于图案化的结构表面，以使异型细胞的共培养和图案化的共培养细胞片的回收用于组织工程中的应用。在水中表现出不同转变温度的温敏聚合物包含聚（N-异丙基丙烯酰胺）（PIPAAm）和作为侧链共接枝到PIPAAm主链上的甲基丙烯酸正丁酯（BMA）。使用电子束聚合方法和多孔金属掩模将se共聚物表面接枝成图案以获得图案化的双重温敏细胞培养表面。在有图案的表面上。大鼠原代肝细胞（HC）和牛颈动脉内皮细胞（EC）的位点选择性粘附和生长允许模式化共培养，利用由培养温度调节的疏水/亲水表面化学作为唯一变量。在27摄氏度。接种的HC仅粘附在疏水的脱水P（IPAAm-BMA）共接枝结构域（1-mm直径区域）上，但不粘附在相邻的水合PIPAAm结构域上。然后，在将培养温度升高至37 ℃时，顺序接种的EC仅粘附于疏水化的PIPAAm结构域，从而实现图案化的共培养。将培养温度降低到20 ℃促进了两种聚合物接枝结构域的水合作用，允许共培养的图案化细胞单层作为具有异型细胞相互作用的连续细胞片层释放。可以操纵、移动和夹在其他结构中的被覆盖的共培养细胞片。为基础细胞生物学研究和通过覆盖共培养的细胞片制备组织模拟多层材料提供了新的有用的构建体。(C)2004 Elsevier Ltd.保留所有权利。
Heterotypic cell interactions are critical to achieve and maintain specific functions in many tissues and organs. We have focused on patterned structure surfaces to enable co-culture of heterotypic cells and recovery of patterned co-cultured cell sheets for applications in tissue engineering. Thermoresponsive polymers exhibiting different transition temperatures in water comprise both poly(N-isopropylacrylamide) (PIPAAm) and n-butyl methacrylate (BMA) co-grafted as side chains to PIPAAm main chains. The se copolymers were surface-grafted in patterns to obtain patterned dual thermoresponsive Cell culture surfaces using electron beam polymerisation method and porous metal masks. On patterned surfaces. site-selective adhesion on and growth of rat primary hepatocytes (HCs) and bovine carotid endothelial cells (ECs) allowed patterned co-culture, exploiting hydrophobic/hydrophilic surface chemistry regulated by culture temperature as the sole variable. At 27degreesC. seeded HCs adhered exclusively onto hydrophobic, dehydrated P(IPAAm-BMA) co-grafted domains (1-mm diamater area), but not onto neighbouring hydrated PIPAAm domains. Sequentially seeded ECs then adhered exclusively to hydrophobised PIPAAm domains upon increasing culture temperature to 37degreesC, achieving patterned co-cultures. Reducing culture temperature to 20degreesC promoted hydration of both polymer-grafted domains, permitting release of the co-cultured, patterned cell monolayers as continuous cell sheets with heterotypic cell interactions. Recovered co-cultured cell sheets can be manipulated, moved and sandwiched with other structures. providing new useful constructs both for basic cell biology research and preparation of tissue-mimicking multi-layer materials through overlaying co-cultured cell sheets. (C) 2004 Elsevier Ltd. All rights reserved.