Hepatic tissue engineering has been investigated for the treatment of liver disease. One promising tissue is the hepatocyte sheet, cultured in temperature-responsive cell culture dishes. However, the preparation of hepatocyte sheets requires precoating of the dish with fetal bovine serum (FBS), which is not desirable for clinical applications. To overcome this issue, we developed a thermoresponsive polymer-modified glass with an affinity for hepatocytes. Poly(--vinylbenzyl----galactopyranosyl-(1→4)--guliconamide)--poly(-isopropylacrylamide) (PVLA--PNIPAAm) was prepared through two steps of atom transfer radical polymerization (ATRP). The prepared copolymer brushes were characterized using X-ray photoelectron spectroscopy, atomic force microscopy, gel permeation chromatography, contact angle measurements, and fluorescence labeled-fibronectin and -lectin adsorption assays. These characterizations showed that the smooth block copolymer layer was successfully modified by ATRP. The feasibility of using the prepared polymer brush-grafted glass as a hepatocyte sheet fabrication dish was investigated using primary rat hepatocytes. The PVLA--PNIPAAm brushes exhibited hepatocyte adhesion and proliferation without FBS precoating. By reducing the temperature from 37 °C to 20 °C, the hepatocyte sheet was recovered from the copolymer brush. These results indicated that the prepared PVLA--PNIPAAm brushes can be utilized for effective hepatocyte sheet fabrication without the need for FBS precoating.

中文翻译：

---

用于有效制造肝细胞片的热响应嵌段共聚物刷修饰界面

肝组织工程已被研究用于治疗肝病。一种有前途的组织是肝细胞片，在温度响应性细胞培养皿中培养。然而，肝细胞片的制备需要用胎牛血清（FBS）预涂培养皿，这对于临床应用来说是不理想的。为了克服这个问题，我们开发了一种对肝细胞具有亲和力的热敏聚合物改性玻璃。通过原子转移自由基聚合(ATRP)两步制备聚(-乙烯基苄基----吡喃半乳糖基-(1→4)--古利康酰胺)-聚(异丙基丙烯酰胺)(PVLA--PNIPAAm)。使用 X 射线光电子能谱、原子力显微镜、凝胶渗透色谱、接触角测量以及荧光标记纤连蛋白和凝集素吸附测定对制备的共聚物刷进行表征。这些表征表明光滑的嵌段共聚物层成功地被 ATRP 改性。使用原代大鼠肝细胞研究了使用制备的聚合物刷接枝玻璃作为肝细胞片制造皿的可行性。 PVLA--PNIPAAm 刷在没有 FBS 预涂的情况下表现出肝细胞粘附和增殖。通过将温度从 37°C 降低至 20°C，从共聚物刷中回收肝细胞片。这些结果表明，制备的 PVLA-PNIPAAm 刷可用于有效的肝细胞片制造，而不需要 FBS 预涂。