Wei Qiao-Ling, Xu Jian-Jiang
Department of Ophthalmology, Eye Ear Nose and Throat Hospital of Fudan University, Shanghai 200031, China.
Zhonghua Yan Ke Za Zhi. 2011 Jan;47(1):75-8.
Cell sheet technology (CST) is based on the use of poly (N-isopropylacrylamide, PNIPAAm), which can be exhibit reversible hydration and dehydration of its polymer chains in response to temperature changes across the lower critical solution temperature(LCST)of 32°C. By reducing the incubation temperature to 20°C, all cultured cells are harvested as intact sheets along with their deposited extracellular matrix (ECM) due to the conversion of the grafted PIPAAm from hydrophobic to hydrophilic, as ECM remains present on the basal surface of the cell sheets, they can maintain cell viability and function as well as directly transplanted to tissue beds or even layered to create three-dimensional (3D) tissue-like structures without any scaffolds or sutures. The temperature-sensitive surfaces' preparation approaches, density, thickness, membrane additive ingredients and so on, all affect cell adhesion and proliferation. It can maintain cell viability and improve function by accelerating cell sheet detachment through changing the membrane compositions, density as well as types of graft substrate. With CST, cultured autologous/allogeneic corneal seed cells in vitro used as transplant sources can overcome the problems of immunorejection of transplanted tissues as well as donor organ shortages. So far, the cell sheet of limbal epithelium and autologous oral mucosal epithelium obtained by the CST have been successfully used in clinical graft for ocular surface reconstruction. Finally, There is an overview of preparations of temperature-responsive surfaces, impacts of various factors that influenced cultured cells in vitro and clinical applications or clinically relevant animal experimentations of CST in corneal tissue engineering.
细胞片技术(CST)基于聚(N-异丙基丙烯酰胺,PNIPAAm)的使用,其聚合物链可在32°C的低临界溶液温度(LCST)范围内响应温度变化而呈现可逆的水合和脱水。通过将孵育温度降至20°C,由于接枝的PIPAAm从疏水性转变为亲水性,所有培养的细胞连同其沉积的细胞外基质(ECM)一起作为完整的薄片收获,因为ECM仍存在于细胞片的基底表面,它们可以维持细胞活力和功能,并且可以直接移植到组织床,甚至可以分层以创建三维(3D)组织样结构,而无需任何支架或缝合。温度敏感表面的制备方法、密度、厚度、膜添加剂成分等都会影响细胞粘附和增殖。通过改变膜组成、密度以及接枝底物的类型来加速细胞片脱离,可维持细胞活力并改善功能。利用CST,体外培养的自体/异体角膜种子细胞作为移植来源可以克服移植组织的免疫排斥问题以及供体器官短缺问题。到目前为止,通过CST获得的角膜缘上皮细胞片和自体口腔黏膜上皮细胞片已成功用于眼表重建的临床移植。最后,综述了温度响应表面的制备、影响体外培养细胞的各种因素的作用以及CST在角膜组织工程中的临床应用或临床相关动物实验。