摘要细胞层片技术(CST)以异内基丙烯酰胺(PNIPAAm)在低临界溶解温度(LCST)快速转变亲水相/疏水相为基础,通过调节细胞的培养温度获取连接紧密的活细胞片与细胞外基质.这种无创性方法获取的活细胞植片,不需支架、不需缝线即可移植,叠加同种或异种细胞层片可构建三维立体组织或器官.温度敏感性材料的制备途径、密度、厚度、膜添加剂成分等都影响着细胞的黏附、增殖;通过改变膜成分与密度、接枝底物种类等,可加速细胞植片脱附,有助于保存细胞活力与功能发挥.以CST为基础,体外培养自体或异体角膜种子细胞为移植源,在降低移植术后排斥与感染同时,还可缓解供体紧张现状,实现一个供体治疗多个患者,为病患提供更多治疗机会,目前通过CST获取的角膜缘上皮细胞及自体口腔黏膜细胞植片已成功应用于临床进行眼表重建.现就温度敏感性材料制备方法及其特点、体外培养细胞影响因素及CST在角膜组织工程研究的应用现状进行阐述.

abstractsCell 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 ℃. By reducing the incubation temperature to 20 ℃, 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 temperatureresponsive 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.