猪主动脉瓣去细胞基质的制备  

Cell extraction from porcine aortic valve

作　　者：顾春虎[1] 王云雅[2] 易定华[1] 张近宝[1] 刘维永[1] 刘洋[1] 

机构地区：[1]第四军医大学西京医院心脏外科,陕西西安710032 [2]第四军医大学西京医院核医学科,陕西西安710032

出　　处：《心脏杂志》2006年第5期528-531,共4页Chinese Heart Journal

基　　金：国家自然科学基金资助项目(No.30271292);陕西省卫生厅基金资助项目(D2045)

摘　　要：目的完全去除猪主动脉瓣细胞,取得良好的去细胞支架材料。方法用5 g/L胰酶对标本中的某些基质和细胞加以消化后,再应用不同去垢剂(十二烷基硫酸钠、脱氧胆脂酸钠、Triton X-100)加以洗脱,结合溶液渗透压改变等去除猪主动脉瓣细胞;标本进行大体、光镜、电镜观察和热皱缩温度的检测。结果胆脂酸钠无法完全脱除猪主动脉瓣细胞,十二烷基硫酸钠和曲拉通均可完全去除猪主动脉瓣细胞,Triton X-100组的主动脉瓣胶原纤维和弹性纤维得以较好保持。结论Triton X-100可以成功脱除猪主动脉瓣细胞,为组织工程瓣膜的研究提供材料。AIM To explore method to remove cellular components from porcine aortic valve, and obtain decellularised scaffolds for tissue engineering of heart valve. METHODS To remove all cells from porcine aortic valve, samples were digested in a 0.1% trypsin solution, followed by multistep detergent-enzymatic process with both hypotonic and hypertonic solutions and ionic detergent. Different detergents （Sodium dodecyl sulfate and sodium deoxycholate, Triton X-100） and enzyme （trypsin） extraction were applied. Specimens were observed grossly and the changes of shrinkage temperature were also studied. Haematoxylin-eosin was performed to confirm the removal of cells and transmission electron microscope was used to observe the integrity of collagen and elastin. RESULTS The cells of porcine aortic valve were removed effectively by Triton X-100 and sodium dodecyl sulfate. Sodium deoxycholate could not effectively remove the cells from the root of aortic valve. The major structural components and shrinkage temperature of decellularised porcine aortic valve by Triton X-100 were well maintained. CONCLUSION Acellular tissue matrix （ACTM） of porcine aortic valve can be successfully obtained through this procedure by Triton X-100.

关 键 词：瓣膜 胰蛋白酶 组织工程 脱细胞基质 

分 类 号：R33[医药卫生—人体生理学] Q813.5[医药卫生—基础医学]