原生质体诱导酶对体外培养的人角膜基质细胞活性的影响  

Toxicity of protoplast induced enzyme on human corneal stromal cells in vitro

作　　者：车成业[1] 牟莹莹[2] 徐强[1] 李娜[1] 贾文妍[1] 李翠[1] 张秋秋[1] 王青[1] 赵桂秋[1] 

机构地区：[1]青岛大学医学院附属医院眼科,中国山东省青岛市266003 [2]潍坊医学院附属医院眼科,中国山东省潍坊市261031

出　　处：《国际眼科杂志》2012年第2期212-214,共3页International Eye Science

基　　金：中国国家自然科学基金资助项目(No.81170825);中国青岛市科技发展计划资助项目[No.11-2-3-1-(5)-nsh;10-3-3-3-10-nsh]~~

摘　　要：目的:检测用以制备烟曲霉菌原生质体的诱导酶对体外培养的人角膜基质细胞活性的影响。方法:将浓度为1g/dL蜗牛酶、1g/dL纤维素酶及0.1g/dL溶壁酶的复合诱导酶液与人角膜基质细胞共同培养15min,4h和8h,采用四氮唑盐代谢法(MTT法)检测不同作用时间的诱导酶对人角膜基质细胞的影响,台盼兰染色法检测诱导酶对人角膜基质细胞存活率的影响与其作用时间的关系。结果:共培养15min及4h后细胞形态未见明显变化,8h后细胞间隙略变小,偶见脱壁漂浮细胞,MTT实验显示诱导酶培养到8h时MTT值仍无明显下降,台盼兰染色显示8h内诱导酶对细胞存活率影响较小。结论:用以制备烟曲霉菌原生质体的诱导酶短时间内对体外培养的人角膜基质细胞活性影响较小,这一浓度的复合诱导酶用于动物模型及人细胞学实验较为安全。AIM：To experiment the toxicity of protoplast induced enzyme on human corneal stromal cells in vitro METHODS：Protoplast induced enzyme included 1g/dL snailase,1g/dL cellulase and 0.1g/dL lysing enzyme.The enzyme and human corneal stromal cells were cultured for 15 minutes,4 hours,8 hours.The effect of enzyme on the human corneal stromal cells was experimented with MTT assay and trypan blue staining.RESULTS：The cell morphology did not change after 15 minutes and 4 hours.The intercellular space of cultured cells was a little smaller after 8 hours,and only to see a few dead cells.MTT value was not noticeable decline after 8 hours.Trypan blue staining showed less impact on cell survival by the enzyme within 8 hours.CONCLUSION：Protoplast induced enzyme showed scarce toxicity on cultured human corneal stromal cell within a short time.The concentration of the enzyme used in future experimental animal models and human cytology experiment is safe.

关 键 词：原生质体 诱导酶 角膜基质细胞 细胞活性 

分 类 号：R772.21[医药卫生—眼科]