两种羊膜脱细胞基质制备方法的比较  

Comparison of the preparative methods of human acellular amniotic membrane matrix

作　　者：雷宁静[1] 胡炜[2] 王成春[1] 朱根明[1] 杨琳琳[1] 关方霞[1] 

机构地区：[1]郑州大学生物工程系,郑州450001 [2]郑州大学第一附属医院神经外科,郑州450052

出　　处：《郑州大学学报（医学版）》2010年第4期572-575,共4页Journal of Zhengzhou University(Medical Sciences)

基　　金：大学生创新性实验计划项目091045923

摘　　要：目的:探究制备羊膜脱细胞基质的方法。方法:取新鲜羊膜标本,分别采用TritonX-100(Triton组)预处理(恒温36h)和甘油预处理(4℃,24h×3次),再经胰蛋白酶、EDTA酶消化(分别用时4h和10h),制备羊膜脱细胞基质。经HE染色和扫描电镜观察2组羊膜脱细胞基质残留细胞数和组织结构,并观察接种的第3代人脐带间质干细胞在2种羊膜脱细胞基质上的黏附、生长状况。结果:上述2种方法制备的羊膜脱细胞基质均为白色、半透明膜状物,无细胞及其他组织残留。Triton组羊膜脱细胞基质结构均匀平整,操作时间短。甘油组样本表面凹凸不平,看不到细胞形态。脐带间质干细胞在2种羊膜脱细胞基质表面黏附、增殖状况均良好,Triton组细胞增殖状况总体优于甘油组(P<0.05)。结论:TritonX-100预处理并酶消化法是制备羊膜脱细胞基质较好的方法。Aim：To compare the two methods for preparing the human acellular amniotic membrane（HAAM） matrix.Methods：The fresh amniotic membrane was collected from puerpera with full term cesarean section and prepared by two methods.Ⅰ：fresh human amnion was placed in TritonX-100 for 36 h,and then treated with trypsin and EDTA for 4 h at 37 ℃.Ⅱ：fresh human amnion was placed in glycerin for 24 h × 3 times,and then treated with trypsin and EDTA for 10 h at 4 ℃.The characters and biocompatibility for umbilical cord derived mesenchymal stem cells on the 2 kinds of HAAM matrix were observed.Results：Both of the two kinds of HAAM matrix were white,semitransparent and flexible,and had no residues of cells.The umbilical cord derived mesenchymal stem cells could attach and grow normally on them,and the proliferation of Triton group was better than that of the glycerin group（P〈0.05）.HAAM matrix prepared by method Ⅰ had even and smooth structure,and spent less time.HAAM matrix prepared by method Ⅱ had accidented surface.Conclusion：Using TritonX-100 to prepare HAAM matrix is better.

关 键 词：羊膜脱细胞基质 脐带间质干细胞 生物相容性 

分 类 号：R318.08[医药卫生—生物医学工程]