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燕山大学课程设计说明书2-丙基戊酸、5-氮杂胞苷对ips细胞诱导率影响的设计学院（系）：环境与化学工程学院年级专业：08级生物化工学号：080110050014学生姓名：陈红指导教师：赵红卫教师职称：讲师燕山大学课程设计（论文）任务书院（系）：环境与化学工程学院基层教学单位：生物工程系学号080110050014学生姓名陈红专业（班级）08级生物化工设计题目2-丙基戊酸、5-氮杂胞苷对ips细胞诱导率的影响设计主要内容1.5-氮杂胞苷对ips细胞诱导率的影响2.2-丙基戊酸对ips细胞诱导率的影响3.2-丙基戊酸、5-氮杂胞苷共同对ips细胞诱导率的影响设计要求1.要有明确设计的目的2.设计合理的操作方式3.设计出合理的测定指标4.设计的内容要与题目基本一致5.设计总结与分析工作量1.至少阅读15篇以上的相关科技文献2.设计文字至少在15000字以上工作计划12.27——12.28查阅资料12.29——12.31整理文献1.1——1.5提出设计方案1.6——1.10撰写说明书1.11——1.12检查内容，准备答辩1.13——1.14答辩参考资料[1]申红芬,姚志芳,肖高芳等.诱导性多潜能干细胞(iPScells)———现状及前景展望[J],生物化学与生物物理进展,2009,36(8):950-960.[2]王孟丽，沈晓丽.体细胞重编程为诱导性多能干细胞的研究进展[J],国外医学.老年医学分册,2009,30(4):50-52.[3]成德,雷蕾,卢智娟等.诱导多能干细胞(iPS)的诱导培养与鉴定[J],生物工程学报,2010,26(4):421-430.[4]苏庸春,徐酉华,蔡倩等.甲基化抑制剂5-氮杂胞苷对HL一60细胞增殖及凋亡的影响[J],医药期刊,2010,35(7):981-984.指导教师签字基层教学单位主任签字说明：学生、指导教师、基层教学单位各一份。年月日燕山大学课程设计成绩评定表设计者姓名:陈红学号:080110050014设计题目:2-丙基戊酸、5-氮杂胞苷对ips细胞诱导率的影响说明书成绩评定（满分100分）：得分设计内容的切题程度：满分20分（）设计内容立题合理性：满分10分（）设计内容的规范程度：满分10分（）设计书前后内容完整：满分20分（）设计说明说排版成绩：满分20分（）设计说明书的工作量：满分10分（）设计过程平时成绩：满分10分（）总分成绩：指导教师：年月日答辩成绩评定：（满分100分）得分仪表成绩：满分20分（）口语表达：满分20分（）幻灯质量：满分20分（）设计分析：满分40分（）总分：教师：年月日设计撰写成绩（70%）答辩成绩（30%）合计教师签字：年月日2010-2011秋季学期生物工程专业课程设计结题论文2-丙基戊酸、5-氮杂胞苷对ips细胞诱导率的影响学院（系）：环境与化学工程学院年级专业：08级生物化工学号：080110050014学生姓名：陈红指导教师：赵红卫教师职称：讲师摘要本设计在用小鼠成纤维细胞诱导ips细胞的培养基中加入小分子化合物2-丙基戊酸、5-氮杂胞苷，探讨它们对ips细胞诱导率的影响。设计分为三部分：5-氮杂胞苷对ips细胞诱导率的影响的设计；2-丙基戊酸对ips细胞诱导率的影响的设计；2-丙基戊酸、5-氮杂胞苷共同对ips细胞诱导率的影响的设计。第一部分设计探索出5-氮杂胞苷对小鼠成纤维细胞转化为ips细胞的诱导率的最适剂量，判断出其影响效果；第二部分设计探索出2-丙基戊酸能使小鼠成纤维细胞转化为ips细胞诱导率最高时的剂量，得出其影响大小；第三部分设计把2-丙基戊酸、5-氮杂胞苷以不同的配比加入培养基中，从而，探索出两种小分子化合物以怎样的比例混合，使ips细胞的诱导率最高。通过本实验设计最终得出2-丙基戊酸、5-氮杂胞苷以及二者的混合物在合适的剂量下哪种方法对ips细胞的诱导率贡献最大，从而得出提高ips细胞诱导率的简易、可行方法。关键词：2-丙基戊酸；5-氮杂胞苷；抑制；甲基化；ips细胞；诱导率I目录第一部分：文献综述1.ips细胞发展简介···········································································11.1ips细胞的诱导·······································································11.1.1ips细胞的诞生·······························································11.1.2导入ips细胞的外源转录因子············································11.1.3受体细胞的选择·····························································21.2ips细胞的表观遗传学······························································31.2.1分子水平上的表观遗传学·················································31.2.2细胞水平上的表观遗传学·················································31.2.3动物水平上的表观遗传学·················································41.3ips细胞的发展及应用······························································41.3.1ips细胞的分化和移植在治疗血液疾病中的作用·····················41.3.2ips细胞对神经系统疾病的治疗作用····································41.3.3ips细胞有望应用于治疗不孕症··········································61.3.4ips细胞可提供体外的疾病模型··········································61.4ips技术方面的障碍·································································72.促进ips细胞诱导率的小分子化合物··················································82.12-丙基戊酸············································································82.25-氮杂胞苷············································································83.总结——本设计研究的价值····························································9第二部分：课程设计部分1.材料·························································································121.1小鼠···················································································121.2细胞株、质粒······································································121.3试验试剂············································································121.4试验器材·············································································122.方法·························································································132.1细胞培养基的制备································································132.1.1MEF培养基的制备·························································132.1.2ES细胞培养基的制备·····················································132.2饲养层细胞的制备································································132.2.1Oct4-EGFP的制备··························································132.2.2MEF饲养层细胞的制备···················································132.3诱导产生ips细胞··································································132.4ips细胞诱导率的侧定·····························································14II2.5ips细胞系的鉴定···································································142.5.1绿色荧光蛋白表达检测···················································142.5.2碱性磷酸酶染色鉴定······················································143.设计·························································································153.15-氮杂胞苷对ips细胞诱导率的影响··········································153.22-丙基戊酸对ips细胞诱导率的影响··········································153.32-丙基戊酸、5-氮杂胞苷共同对ips细胞诱导率的影响···················154.分析与总结················································································155.课程设计心得············································································16参考文献·······················································································18燕山大学课程设计说明书1第一部分文献综述1．ips细胞发展简介1.1ips细胞的诱导1.1.1i