绵羊颈动脉脱细胞基质制备方法的研究.pdfVIP

中国农业科学 2007,40(12):2920-2925 Scientia Agricultura Sinica 绵羊颈动脉脱细胞基质制备方法的研究 张翊华，窦忠英 （西北农林科技大学/ 陕西省干细胞工程技术研究中心，杨凌 712100 ） 摘要：【目的】证明酶-去污剂法制备绵羊颈动脉脱细胞基质的可行性，进一步探索制备绵羊颈动脉脱细胞基 质的新方法，制备比较理想的绵羊颈动脉脱细胞基质，为构建组织工程化血管提供支架材料。【方法】分别采用反 复冻融-1%曲拉通（Triton X-100）加蛋白酶抑制剂（PMSF）法（蛋白酶抑制剂（PMSF）-1%曲拉通（Triton X-100） 反复冻融法）和0.25%胰蛋白酶-1%Triton X-100法制备绵羊颈动脉脱细胞基质，标本进行大体、光镜和扫描电镜 观察，力学性能检测并做比较。【结果】反复冻融-1%Triton X-100加PMSF法（PMSF-1%Triton X反复冻融法）不 仅能完全脱除细胞、保持基质纤维的正常结构，而且制备的脱细胞基质能保持良好的形状和力学性能、不塌陷； 0.25%胰蛋白酶-1%Triton X-100法虽能完全脱除细胞，但制备的脱细胞基质不能保持良好的形状和张力强度，有 不同程度的管腔塌陷。【结论】与0.25%胰蛋白酶-1%Triton X-100法比较，反复冻融-1%Triton X-100加PMSF法 是制备绵羊颈动脉脱细胞基质更好的新方法。 关键词：颈动脉；脱细胞基质；绵羊 Study on Methods for Preparing Acellular Matrices of Sheep Carotids ZHANG Yi-hua, DOU Zhong-ying (The Research Centre of Stem Cell Engineering Technique in Shaanxi Province, Northwest Sci-Tech University of Agriculture and Forestry, Yangling 712100) Abstract: 【Objective 】This paper is to test the feasibility of preparing acellular matrices of sheep carotids with enzyme-eradicator method, further explore a new method, and prepare good acellular matrices of sheep carotids to provide scaffolds for constructing tissue-engineering blood vessels. 【Method 】Methods A and B were used for preparing acellular matrices of sheep carotids. A method was that sheep carotids (n=20) were frozen and thawed repeatedly for four times, then treated with phosphate-buffered saline (PBS) containing 1%Triton X-100 and 1μmol·L-1 phenyl methyl sulfonyl fluoride (PMSF) for 4-7 d. B method was that sheep carotids (n=20) were treated with PBS containing 0.25% trypsin and 0.04% ethylene diamine tetraacetic acid ( EDTA) for 1 d, and then with