von Recum H, Kikuchi A, Okuhara M, Sakurai Y, Okano T, Kim S W
Department of Bioengineering, University of Utah, Salt Lake City 84112, USA.
J Biomater Sci Polym Ed. 1998;9(11):1241-53. doi: 10.1163/156856298x00758.
A cross-linkable co-polymer of UV-sensitive 4-(N-cinnamoylcarbamide)methylstyrene (CCMS) and N-isopropylacrylamide (NIPAAm), was applied to porous tissue culture inserts. Surface chemical analyses of the inserts show an introduction of a thermally responsive polymer comparable to that on similarly incorporated non-porous polystyrene surfaces. Contact angle measurements as well as atomic force microscopy show a surface change in response to changing temperature in an aqueous environment, from hydrophilic, extended polymer chains below 32 degrees C to a dense hydrophobic film above 32 degrees C. Cell growth on porous inserts allowed measurement of cell expression, such as transepithelial resistance and fluid transport, which are not observable on cells from non-porous surfaces. Cultures of retinal pigmented epithelium (RPE) were able to restore an environment similar to in vivo by forming a tight junction barrier membrane upon confluence at 37 degrees C, as observed by changes in morphology, transepithelial resistance, and directionally-specific fluid transport. In addition, cells cultured on these surfaces detached as an oriented polarized sheet when the inserts were brought to 20 degrees C. This cell sheet was transplanted to other tissue culture surface without polymer detachment or dissolution, or cell damage caused by traditional detachment methods using proteolytic enzymes.
一种由对紫外线敏感的4-(N-肉桂酰氨基甲酰)甲基苯乙烯(CCMS)和N-异丙基丙烯酰胺(NIPAAm)组成的可交联共聚物被应用于多孔组织培养插入物。对插入物的表面化学分析表明，引入了一种热响应聚合物，其与类似掺入的无孔聚苯乙烯表面上的聚合物相当。接触角测量以及原子力显微镜显示，在水性环境中，随着温度变化，表面会发生变化，从32℃以下的亲水性、伸展的聚合物链变为32℃以上的致密疏水膜。多孔插入物上的细胞生长使得能够测量细胞表达，如跨上皮电阻和液体运输，这在无孔表面的细胞上是无法观察到的。视网膜色素上皮(RPE)培养物在37℃汇合时能够通过形成紧密连接屏障膜来恢复类似于体内的环境，这可以通过形态学、跨上皮电阻和定向特异性液体运输的变化观察到。此外，当插入物温度降至20℃时，在这些表面上培养的细胞会作为一个定向极化片脱落。这个细胞片被移植到其他组织培养表面，没有聚合物脱落或溶解，也没有因使用蛋白水解酶的传统脱落方法而导致细胞损伤。
