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张 岚,舒 峻,蔡 哲,付桂香,徐 杨,高 艳,崔晓惠.人羊膜上皮细胞分离的正交法研究[J].中国康复医学杂志,2009,24(4):352~354
人羊膜上皮细胞分离的正交法研究    点此下载全文
张 岚  舒 峻  蔡 哲  付桂香  徐 杨  高 艳  崔晓惠
中日友好医院临床医学研究所,北京,100029
基金项目:北京市自然科学基金重点项目(5041002),北京市科委重大研究课题(D07050701350704),国家自然科学基金(30471773)
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摘要:
      目的:研究人羊膜上皮细胞(hAECs)分离的适宜条件。方法:采用正交试验的方法,以L9(34)正交表安排影响hAECs分离的几个主要因素,每组实验所得细胞计数并培养24h后,收集悬浮细胞计算细胞贴壁率,培养7d计算扩增细胞数,以此为指标分析各因素对hAECs分离的影响。结果:胰蛋白酶的浓度及消化次数对细胞得量均有显著影响(P<0.05)。结论:应用正交试验法得出hAECs分离的适宜条件为:胰蛋白酶液与羊膜组织体积比为1∶1,胰蛋白酶浓度0.25%,消化时间10min/time,消化4次。
关键词:人羊膜  上皮细胞  分离方法  正交试验法
Orthogonal research on separation of human fetus amnion epithelial cells    Download Fulltext
Institute of Clinical-Medicine Science China-Japan Friendship Hospital, Beijing, 100029
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Abstract:
      Objective: To study the appropriate conditions of separation of human fetus amnion epithelial cells (hAECs). Method: The orthogonal test was used to arrange those several major factors which impact the separation of hAECs as L9(34) orthogonal table. After counting numbers, the hAECs from different test groups were cultivated for 24h, then collected the suspended cells and calculated the adherence rate. The number of proliferated cells were counted after 7 days, and all these data were used to analyze the affection of the factors on the hAECs separation. Result: The concentration of trypsin and the times of trypsinization were the key factors which impacted on the hAECs separation(P<0.05). Conclusion: The appropriate conditions of the hAECs separation are as follows: the volume of 0.25% trypsin is equal to the amnion;trypsinizing four times,10 min/time.
Keywords:human amnion  epithelial cells  separation  orthogonal test
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