中文摘要：

研究了温度梯度法所制再生丝素蛋白膜的溶失率、抗酶能力和动物细胞体外培养特性。以紫外分光光度法检测丝素蛋白膜在水中的溶失率;再用质量体积浓度0.25%和0.5%的胰蛋白酶液分别消化丝素蛋白膜2d,以膜的结构和质量是否发生明显变化为指标,判断膜抗胰蛋白酶的能力;最后在丝素蛋白膜上培养BHK-21细胞和Vero细胞,再用0.25%和0.5%胰蛋白酶液分别消化膜上细胞并进行传代培养,观察细胞形态和生长状况。实验结果表明,温度梯度法直接所制丝素蛋白膜48h内的溶失率低于0.2%,再经25%甲醇处理后溶失率为0,2d内膜仍保持完整结构且质量无明显变化;BHK-21细胞和Vero细胞在丝素蛋白膜上生长良好,形态正常;0.25%和0.5%的胰蛋白酶液均可在4min内将膜上两种细胞消化脱落且细胞连续传代培养5代,细胞形态正常。

英文摘要：

The dissolve-loss rate and antitrypsin ability of regenerated silk fibroin membrane and its characteristics as animal cells culture in vitro were studied by temperature gradient method.Dissolve-loss rate of the fibroin membrane in water by ultraviolet spectrophotometry was detected.The concentration of 0.25%and 0.5%trypsin was used respectively to digest regenerated silk fibroin membrane for 48 hto test its structure and judge its antitrypsin ability according to structure and weight change.Silk fibroin membrane as matrix was implemented to culture BHK-21 and Vero cells to observe their morphology and trypsin digestion characteristics.However,the results show that dissolve-loss rate of silk fibroin membranes was less than 0.2%and was 0treated with 25% methanol within 48 hin water.The structure of regenerated silk fibroin membrane remained completely integration treating for 48 husing 0.25%and 0.5%trypsin solution and its weight had no obvious change.BHK-21 cells and Vero cell were grown well using silk fibroin membrane as matrix in vitro.Specifically,the digest time of anchorage-dependent BHK-21 cells and Vero cells using 0.25% and 0.5%trypsin solution were both within 4min,keeping normal cell morphology,and the digested cells remained normal growth for 5generations in T-Flask culture.