再生丝素蛋白膜的性能及其细胞培养特性研究  

Study on Properties of Regenerated Silk Fibroin Membrane and Its Characteristics as Cell Culture Matrix

作　　者：龙仕和[1,2] 李明生[1,3] 靳冬武 崔梦楠[2] 马素娟[4] 徐水林[1,3] 柏家林[1,2] 李倬[1,2] 冯玉萍[1,2] 马忠仁[1] 

机构地区：[1]西北民族大学生物工程与技术国家民委重点实验室,兰州730030 [2]西北民族大学生命科学与工程学院,兰州730030 [3]甘肃省动物细胞工程技术研究中心,兰州730030 [4]兰州理工大学生命科学与工程学院,兰州730050

出　　处：《材料导报》2015年第6期65-70,共6页Materials Reports

基　　金：教育部"创新团队发展计划"(IRT13091);校中青年科研基金(X2006-023);西北民族大学中央专项资金资助研究生项目(ycx14033);中央高校项目(zyz2012088)

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。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.

关 键 词：再生丝素蛋白膜 溶失率 消化 细胞培养基质 

分 类 号：Q813[生物学—生物工程] TB34[一般工业技术—材料科学与工程]