动物细胞培养——基本技术和特殊应用指南（原书第7版）

• 萌宠菠菠乐园
• 2024-10-23 05:03

ID:12659339,SiteID:122,InstanceID:17739,Title:动物细胞培养——基本技术和特殊应用指南（原书第7版）,TitleAbbrev:,SubTitle:,ETitle:Culture of Animal Cells: A Manual of Basic Technique and Specialized Applications（7th Edition）,ETitleAbbrev:,ESubTitle:,Fileref:/web/resource/import/xml/importxml/9787030603593/XML/9787030603593.xml,Annotation:null,AbstractCH:自本书第6版发行以来，细胞培养领域已取得诸多进展，第7版除保留基本内容（培养细胞的生物学、细胞培养实验室的设计与布局、培养器皿与培养基、原代培养、细胞系及其鉴定、污染、分化、老化、永生化等）之外，特别强化了某些章节的内容，尤其见于3D培养、干细胞、特殊类型细胞培养、STR测序、规模化培养等的介绍，因此第7版无论是在理论上还是在技术上均更具新颖性和实用性。本书可供学术研究人员、临床医生、生物制药研究者、有关学科大学师生尤其是研究生，以及细胞和分子生物学与遗传学研究及实验室管理人员，特别是培训老师、技术人员与受培训人员阅读参考。,AbstractEN:,ZhKeyWords:319,151,EnKeyWords:,Height:null,Width:null,Printing:null,Pheiclasscode:null,EpubID:0,MobiID:0,PublishID:0,TxtID:0,HighPdfID:0,PdfID:0,LogoID:12659340,BookCoverPath:null,BookType:7,BookOtherType:null,Volumenum:0,Cip:null,Releaseinfo:,CheckDate:null,CopyRightBeginDate:null,CopyrightYear:null,Allrightreserved:null,RegistrationDate:null,DeriesID:3172,SeriesName:生命科学实验指南系列null,EbookPrice:0.0,EntityPrice:328.00,Status:6,Flag:null,ISBN:978-7-03-060359-3,XmlID:B1F1A5279D2DC406E898E48C19D45063E000,Language:,legalnotice:null,Themeword:,PublishArea:null,Publisher:科学出版社,PrintNum:1,Type:null,PublishDate:2019-01-01 00:00:00.0,UpdateDate:null,Size:null,Format:,PrintCompany:null,PrintBind:null,Foldout:null,Words:1185.0,Sheets:null,YinCi:null,Impressions:null,Foundation:null,PaperCount:null,MaxPageNO:840,PdfTotalCount:0,HitCount:0,TopMark:0,OrderFlag:0,Recommend:null,PublishTimes:1,AuthorInfos:{"R.Ian Freshney":{"Role":"","Blurb":null},"章静波":{"Role":"","Blurb":null},"徐存拴":{"Role":"","Blurb":null},"李悦":{"Role":"","Blurb":null},"刘晶":{"Role":"","Blurb":null}},IsConvert:N,IndexStatus:null,Prop1:null,Prop2:null,Prop3:null,Prop4:null,AddUser:kongrongjie,AddTime:2022-11-30 14:29:53.0,ModifyUser:liudong,ModifyTime:2024-07-08 09:27:06.0,Weight:null,IsProvideDown:null,ImportResourceFormat:null,Href:,OldID:0,SeriesAlias:null,InnerCode:生命科学实验指南系列,SeriesIndex:null,RefCount:0,EquationCount:0,FigCount:305,TableCount:123,Authors:R.Ian 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简介目录文章

自本书第6版发行以来，细胞培养领域已取得诸多进展，第7版除保留基本内容（培养细胞的生物学、细胞培养实验室的设计与布局、培养器皿与培养基、原代培养、细胞系及其鉴定、污染、分化、老化、永生化等）之外，特别强化了某些章节的内容，尤其见于3D培养、干细胞、特殊类型细胞培养、STR测序、规模化培养等的介绍，因此第7版无论是在理论上还是在技术上均更具新颖性和实用性。本书可供学术研究人员、临床医生、生物制药研究者、有关学科大学师生尤其是研究生，以及细胞和分子生物学与遗传学研究及实验室管理人员，特别是培训老师、技术人员与受培训人员阅读参考。

参译人员（以章节排序）译者的话前言和致谢缩略语第1章 绪论1.1 历史背景1.2 组织培养的优点1.2.1 环境控制1.2.2 样品的特征和均一性1.2.3 经济、规模和机械化1.2.4 体内环境的体外模拟1.3 局限性1.3.1 专业技能1.3.2 量的问题1.3.3 去分化和选择1.3.4 细胞的起源1.3.5 不稳定性1.4 体外的主要差异1.5 组织培养的类型第2章 培养细胞的生物学2.1 细胞培养环境2.2 细胞黏附2.2.1 细胞黏附分子2.2.2 细胞间连接2.2.3 细胞骨架2.2.4 细胞外基质2.2.5 细胞迁移2.3 细胞增殖2.3.1 细胞周期2.3.2 细胞增殖调控2.4 细胞分化2.4.1 细胞分化的诱导和保持2.4.2 细胞分化及去分化潜能2.5 细胞信号转导2.6 能量代谢2.7 培养细胞的起源2.7.1 培养的开始2.7.2 细胞系的演化2.7.3 细胞的衰老2.7.4 连续细胞系的转化和建立2.8 术语定义第3章 实验室设计及布局3.1 布局、陈设和服务设施3.1.1 必备条件3.1.2 服务3.1.3 通风装置3.2 设计与布局3.2.1 无菌操作区3.2.2 层流柜3.2.3 工作台3.2.4 检疫和防范3.2.5 培养3.2.6 准备区3.2.7 储存3.3 灾害管理第4章 设备和材料4.1 组织培养实验室的要求4.2 无菌工作区4.2.1 层流生物安全柜4.2.2 手推车4.2.3 无菌液体处理——移液和分液4.2.4 倒置显微镜4.2.5 照相机和监视器4.2.6 解剖显微镜4.2.7 离心机4.2.8 细胞计数4.3 孵育和培养4.3.1 恒温箱4.3.2 湿式CO2培养箱4.3.3 温度记录仪4.3.4 滚筒支架4.3.5 磁力搅拌器4.3.6 培养器皿4.4 准备和灭菌4.4.1 洗涤4.4.2 培养基和试剂的制备4.4.3 灭菌4.5 储存4.5.1 耗材4.5.2 冰箱和冷冻箱4.5.3 冻存容器4.5.4 程序降温仪4.6 实验室辅助设备4.6.1 计算机和网络4.6.2 正置显微镜4.6.3 低温冷冻箱4.6.4 共聚焦显微镜4.6.5 PCR热循环仪4.7 专用设备4.7.1 显微注射装置4.7.2 菌落计数器4.7.3 离心式淘洗器4.7.4 流式细胞仪第5章 无菌技术5.1 无菌技术的目的5.1.1 污染风险5.1.2 维持无菌状态5.2 无菌环境的基本要素5.2.1 层流5.2.2 安静区域5.2.3 操作台5.2.4 个人卫生5.2.5 试剂和培养基5.2.6 培养物5.3 无菌处理5.3.1 擦拭5.3.2 加盖5.3.3 灼烧5.3.4 试剂瓶和培养瓶的操作5.3.5 移液5.3.6 大容量移液5.3.7 液体倾倒5.4 标准化操作5.4.1 培养瓶和试剂瓶5.4.2 培养皿和多孔培养板5.5 仪器和设备5.5.1 冰箱和冷室5.5.2 恒温箱5.5.3 盒装培养物5.5.4 充入CO2气体第6章 安全性、生物伦理及验证6.1 实验室安全6.2 危险性评估6.3 标准操作程序6.4 安全规则6.5 常规安全6.5.1 操作者6.5.2 设备6.5.3 玻璃器皿和锐利物品6.5.4 化学毒性6.5.5 气体6.5.6 液氮6.5.7 烧伤6.6 火6.7 电离辐射6.7.1 摄入6.7.2 放射性废物的处理6.7.3 标记试剂的辐照6.7.4 高能源辐照6.8 生物危害性6.8.1 生物防范水平6.8.2 生物安全柜（BSC）6.8.3 人体活检材料6.8.4 细胞系6.8.5 遗传操作6.8.6 生物危险性废物的处理6.8.7 净化和熏蒸6.9 生物伦理6.9.1 动物组织6.9.2 人体组织6.10 质量保证6.10.1 操作程序6.10.2 质量控制（QC）6.10.3 确认6.10.4 身份验证6.10.5 起源6.10.6 污染第7章 培养器皿和附着物7.1 附着物7.1.1 附着和生长7.1.2 常用的附着材料7.1.3 附着物替代品7.2 表面处理7.2.1 附着物包被7.2.2 饲养层7.2.3 非黏性附着物7.3 培养器皿的选择7.3.1 细胞产量7.3.2 多孔板7.3.3 培养瓶和培养皿7.3.4 高细胞产量7.3.5 悬浮培养7.3.6 通气7.3.7 取样和分析7.3.8 不均匀生长7.3.9 成本7.4 特殊系统7.4.1 渗透性支持物7.4.2 三维基质第8章 成分明确培养基及补充物8.1 培养基的发展史8.2 物理化学特征8.2.1 pH8.2.2 CO2和碳酸盐8.2.3 缓冲作用8.2.4 氧气8.2.5 渗透压8.2.6 温度8.2.7 黏度8.2.8 表面张力和成泡性8.3 平衡盐溶液8.4 完全培养基8.4.1 氨基酸8.4.2 维生素8.4.3 盐类8.4.4 葡萄糖8.4.5 有机补充物8.4.6 激素和生长因子8.4.7 抗生素8.5 血清8.5.1 蛋白质8.5.2 生长因子8.5.3 激素8.5.4 营养物及代谢物8.5.5 脂类8.5.6 矿物质8.5.7 抑制物8.6 培养基和血清的选择8.6.1 血清批次预约8.6.2 血清的测试8.6.3 热灭活8.7 其他添加物8.7.1 氨基酸水解物8.7.2 胚胎浸出液8.7.3 适应性培养基8.8 储存第9章 无血清培养基9.1 血清的缺点9.2 无血清培养基的优点9.3 无血清培养基的缺点9.4 血清的替代9.4.1 无血清培养基的商业供应9.4.2 血清替代物9.4.3 无血清传代培养9.4.4 激素9.4.5 生长因子9.4.6 血清中的营养物质9.4.7 蛋白质和多聚胺9.4.8 黏度9.5 无血清培养基的研发9.6 无血清培养基的选择9.6.1 细胞或产物特异性9.6.2 细胞系对无血清培养基的适应性9.7 无血清培养基的制备9.8 无动物蛋白培养基9.9 小结第10章 准备与灭菌10.1 准备试剂与用品10.2 设备与液体的灭菌10.2.1 干热灭菌10.2.2 压力-蒸汽灭菌10.2.3 射线灭菌10.2.4 化学灭菌10.2.5 无菌指示器10.2.6 过滤除菌10.3 设备10.3.1 玻璃器皿10.3.2 玻璃移液管10.3.3 螺口盖10.3.4 清洁剂的选择10.3.5 种类繁杂的设备10.3.6 可重复使用的灭菌过滤器10.4 试剂与培养基10.4.1 水10.4.2 纯水器的维护10.4.3 平衡盐溶液10.4.4 培养基的配制与灭菌10.5 培养基的灭菌10.5.1 可高压灭菌的培养基10.5.2 除菌过滤10.5.3 血清10.5.4 其他试剂的准备与灭菌10.6 培养基的质控、检测和储存10.6.1 质量控制10.6.2 无菌检测10.6.3 培养基和血清的培养检测10.6.4 储存第11章 原代培养11.1 原代培养入门11.1.1 用于解离组织的蛋白酶11.1.2 解离过程的共同特点11.2 组织分离11.2.1 小鼠胚胎11.2.2 鸡胚11.2.3 人体活检材料11.3 原代培养的类型11.3.1 原代外植11.3.2 酶的解离作用11.3.3 温胰蛋白酶消化11.3.4 低温预处理的胰蛋白酶消化11.3.5 鸡胚器官原基11.3.6 其他酶解过程11.3.7 胶原酶11.3.8 机械法解离11.3.9 分离活细胞11.3.10 原代培养小结11.3.11 原始记录第12章 传代培养和细胞系12.1 术语定义12.2 传代培养和扩增12.2.1 交叉污染和鉴定错误12.2.2 支原体污染12.2.3 细胞系的命名12.2.4 培养物的年龄12.2.5 有限细胞系和连续细胞系12.3 选择细胞系12.4 常规培养12.4.1 细胞形态的意义12.4.2 培养基的更换12.4.3 标准的换液方案12.5 传代12.5.1 传代标准12.5.2 单层生长细胞典型的传代方案12.5.3 生长周期和传代比率12.5.4 悬浮培养细胞的扩增12.5.5 悬浮生长细胞的传代12.5.6 培养条件的标准化12.5.7 抗生素的使用12.5.8 培养记录第13章 身份认证和验证13.1 细胞系的身份认证13.1.1 错误身份认定和交叉污染13.1.2 错误身份认定和交叉污染的原因13.1.3 避免错误身份认定和交叉污染13.1.4 细胞系身份认证的技术13.1.5 DNA绘谱13.1.6 通过DNA条码测定动物细胞的种属13.1.7 同工酶分析13.1.8 染色体含量13.1.9 染色体显带技术13.1.10 染色体分析13.1.11 身份认证的必要性13.2 细胞身份的验证13.2.1 细胞的身世记录13.2.2 微生物污染13.2.3 步骤13.3 质量保证13.3.1 细胞系13.3.2 培养基和试剂13.3.3 培养器皿13.3.4 仪器13.3.5 设施第14章 污染14.1 污染的来源14.1.1 操作者的技术14.1.2 环境14.1.3 生物安全柜的使用和维护14.1.4 湿度恒温箱14.1.5 冷藏库14.1.6 无菌材料14.1.7 引入的细胞系和活组织14.1.8 隔离14.2 微生物污染的种类14.3 污染物的监控14.3.1 可见的微生物污染14.3.2 支原体14.3.3 支原体的荧光染色14.3.4 PCR法检测支原体14.3.5 检测支原体的替代方法14.3.6 支原体检测服务14.3.7 病毒污染14.4 污染培养物的处理14.5 污染的去除14.5.1 细菌、真菌和酵母菌14.5.2 支原体的消除14.5.3 清除病毒污染14.5.4 顽固污染14.6 交叉污染14.7 结论第15章 冻存和建库15.1 冻存的意义15.2 冻存前的注意事项15.2.1 鉴定15.2.2 何时冻存15.3 冻存细则15.3.1 细胞冻存的理论背景15.3.2 细胞浓度15.3.3 冻存液15.3.4 冷却速度15.3.5 冻存管15.3.6 低温冰箱15.3.7 培养细胞的冻存15.3.8　冻存记录15.3.9 冻存管的解冻15.3.10 培养瓶冻存15.3.11 玻璃化保存15.4 冻存库的设计和监控15.4.1 库存管理、分配和使用15.4.2 细胞库存的连续更换15.5 细胞库15.6 细胞的运输15.6.1 冻存管15.6.2 活细胞第16章 克隆培养及筛选16.1 细胞的克隆培养16.2 贴壁率的刺激16.2.1 促进克隆生长的条件16.2.2 条件培养基16.2.3 饲养层细胞16.3 悬浮克隆培养16.4 细胞克隆的分离16.4.1 单层贴壁细胞克隆的其他分离技术16.4.2 悬浮细胞克隆16.5 复制性培养16.6 选择性培养基和抑制剂16.7 细胞与基质的相互作用16.7.1 选择性黏附16.7.2 选择性脱壁16.7.3 基质性质16.7.4 选择性饲养层16.7.5 半固体培养基选择第17章 细胞分选17.1 细胞密度及等密度沉降法17.2 细胞体积与沉降速率17.2.1 单位重力沉降17.2.2 离心淘洗分离技术17.3 以抗体为基础的分选技术17.3.1 免疫磁珠分选17.3.2 荧光活化的细胞分选法17.4 初试细胞分离者的选择第18章 细胞系鉴定18.1 鉴定的需求18.2 基因型鉴定18.2.1 真实性验证18.2.2 种属鉴定18.2.3 转化18.3 表型鉴定18.3.1 谱系或组织标记18.3.2 独特标记物18.4 细胞形态学18.4.1 显微镜18.4.2 染色18.4.3 细胞学检查所用培养器皿：单层培养18.4.4 悬浮细胞细胞学检查的标本制备18.4.5 光学显微照相技术18.4.6 活细胞成像18.4.7 共聚焦显微镜18.5 抗原标记18.5.1 免疫染色18.5.2 免疫分析18.6 间隔性记录18.7 细胞周期18.7.1 细胞分离18.7.2 阻断第19章 分化19.1 体内表型的表达19.1.1 去分化19.1.2 细胞谱系选择19.2 分化阶段19.3 增殖和分化19.4 定向和细胞谱系19.5 干细胞可塑性19.6 分化标记物19.7 诱导分化19.7.1 细胞相互作用19.7.2 全身性因子19.7.3 细胞-基质相互作用19.7.4 极性和细胞形状19.7.5 动态应激19.7.6 氧张力19.8 分化和恶性19.9 应用第20章 三维培养20.1 细胞的相互作用和表型表达20.1.1 三维培养的优点和局限性20.1.2 模型的选择20.2 器官培养20.2.1 气体和营养物质的交换20.2.2 结构的完整性20.2.3 生长与分化20.2.4 器官培养的局限性20.2.5 器官培养的类型20.3 组织型培养20.3.1 凝胶和海绵技术20.3.2 中空纤维20.3.3 细胞球体20.3.4 悬滴培养20.3.5 微载体20.3.6 类器官20.3.7 转动室系统20.3.8 藻酸盐活细胞固定术20.3.9 滤膜小皿插件20.3.10 神经聚集物的培养20.4 器官型培养20.4.1 组织等同物20.4.2 组织工程20.5 三维构建物中细胞的成像第21章 规模与自动化21.1 规模悬浮培养21.1.1 连续和分批培养21.1.2 一次性生物反应器21.1.3 规模和复杂性21.1.4 搅匀和换气21.2 单层规模培养21.2.1 多表面扩增器21.2.2 旋转培养21.2.3 微载体21.2.4 巨型微载体21.2.5 灌流式单层培养21.3 程序控制21.4 大规模培养程序21.5 自动化21.5.1 机器人培养细胞21.5.2 高通量筛选（HTS）21.5.3 三维高通量筛选第22章 衰老，永生化和转化22.1 在细胞系特征描述中的作用22.2 遗传不稳定性和异质性22.2.1 遗传不稳定性22.2.2 染色体畸变22.3 永生化22.3.1 衰老的控制22.3.2 病毒基因致永生化22.3.3 人成纤维细胞的永生化22.3.4 端粒酶诱导的永生化22.3.5 供选择的永生化策略22.4 生长控制异常22.4.1 停泊不依赖性22.4.2 接触抑制22.4.3 血清依赖22.4.4 癌基因22.5 致瘤性22.5.1 恶性化22.5.2 肿瘤移植22.5.3 侵袭力22.5.4 血管发生22.5.5 纤溶酶原活化因子第23章 定量23.1 细胞计数23.1.1 血细胞计数器23.1.2 电子计数23.1.3 染色的单层细胞23.1.4 流式细胞仪23.2 细胞重量和细胞压积23.3 DNA含量23.4 蛋白质23.5 合成速率23.5.1 DNA合成23.5.2 蛋白质合成23.6 准备样品用于酶分析和免疫分析23.7 细胞计数23.7.1 原位标记23.7.2 流式细胞术23.8 重复取样23.8.1 数据获得23.8.2 数据分析23.9 细胞增殖23.9.1 实验设计23.9.2 生长周期23.9.3 单层细胞生长曲线的分析23.9.4 培养基体积、细胞浓度和细胞密度23.9.5 悬浮培养23.9.6 生长周期的各个阶段23.9.7 生长曲线的衍生物23.10 贴壁效率23.10.1 集落形成分析23.10.2 自动集落计数23.11 标记指数23.11.1 生长分数23.11.2 有丝分裂指数23.11.3 分裂指数23.12 放射自显影术23.13 细胞周期时间23.14 细胞迁移第24章 细胞毒性24.1 活性、毒性和存活24.2 体外局限性24.2.1 药物代谢动力学24.2.2 新陈代谢24.2.3 组织和全身反应24.3 分析方法的类型24.3.1 生存力24.3.2 存活24.3.3 细胞增殖测定24.3.4 代谢性细胞毒性测定24.3.5 微滴定实验24.3.6 微滴定与克隆存活的比较24.3.7 药物的相互作用24.4 细胞毒性实验的应用24.4.1 抗癌药物筛选24.4.2 肿瘤药物的前瞻性实验24.4.3 药物学实验24.5 基因毒性24.5.1 姐妹染色单体交换的诱变实验24.5.2 致癌性24.6 炎症反应第25章 特殊类型细胞的培养25.1 特殊细胞培养技术25.2 上皮细胞25.2.1 表皮25.2.2 角膜25.2.3 乳腺25.2.4 子宫颈25.2.5 胃肠道25.2.6 肝25.2.7 人肝细胞系HepaRG25.2.8 胰25.2.9 气管和支气管上皮25.2.10 口腔上皮25.2.11 前列腺25.3 间充质细胞25.3.1 结缔组织25.3.2 脂肪组织25.3.3 肌25.3.4 软骨25.3.5 骨25.3.6 内皮细胞25.4 神经外胚层细胞25.4.1 神经细胞25.4.2 神经胶质细胞25.4.3 内分泌细胞25.4.4 黑素细胞25.5 造血细胞25.5.1 红细胞25.5.2 髓细胞和巨噬细胞25.5.3 淋巴细胞25.6 单克隆抗体的制备25.7 生殖腺25.7.1 卵巢25.7.2 睾丸25.8 冷血动物细胞的培养25.8.1 鱼细胞25.8.2 昆虫细胞25.9 肿瘤细胞培养25.10 取材25.10.1 代表性细胞的选择25.10.2 组织的冷冻保存25.11 分离25.12 原代培养25.13 肿瘤细胞的选择培养25.13.1 选择培养基25.13.2 汇合饲养层25.13.3 悬浮克隆25.13.4 异种移植25.14 细胞系的建立25.14.1 原代肿瘤培养物的传代25.14.2 连续细胞系25.15 肿瘤细胞培养物的特征25.15.1 肿瘤细胞的异质性25.15.2 组织型培养25.16 特殊类型肿瘤25.16.1 乳腺25.16.2 肺25.16.3 胃25.16.4 结肠25.16.5 胰腺25.16.6 卵巢25.16.7 前列腺25.16.8 膀胱25.16.9 皮肤25.16.10 子宫颈25.16.11 胶质细胞瘤25.16.12 神经母细胞瘤25.16.13 精原细胞瘤25.16.14 淋巴瘤和白血病第26章 干细胞26.1 胚胎干细胞26.1.1 小鼠胚胎干细胞的诱导26.1.2 小鼠胚胎干细胞的传代培养和扩增26.1.3 人胚胎干细胞的原代培养26.1.4 鱼胚胎来源的多能干细胞26.2 生殖细胞26.3 胚外细胞26.3.1 羊水细胞的培养26.3.2 从新生儿或青年来源的细胞26.3.3 成人来源的多能干细胞26.3.4 人骨髓来源间充质干细胞26.3.5 前列腺上皮干细胞26.3.6 牙上皮干细胞26.3.7 诱导多潜能干细胞26.4 造血干细胞26.4.1 小鼠骨髓长期培养26.4.2 人骨髓长期培养26.5 再生医学中干细胞的应用26.6 肿瘤干细胞第27章 培训纲要27.1 目的27.2 实验制备及操作技巧27.3 基本的细胞培养技术27.4 高阶练习27.5 特殊练习第28章 问题与对策28.1 细胞异常形态28.2 细胞生长缓慢28.2.1 仅限于自己细胞出了问题28.2.2 其他人也遇到同样的问题28.3 培养基28.3.1 试剂配方、准备和存储28.3.2 不稳定试剂28.3.3 配制培养基各种成分的纯度28.4 基质和容器28.5 微生物污染28.5.1 仅限于单个实验者的问题28.5.2 普遍问题28.5.3 室内供气和层流净化工作台28.5.4 特定污染28.6 化学污染28.6.1 玻璃器皿28.6.2 移液管28.6.3 水的纯化28.6.4 低温冻存28.6.5 粉末或气溶胶28.7 原代培养28.7.1 原代培养的存活率低28.7.2 选择细胞错误28.7.3 污染28.8 分化28.9 饲养28.9.1 常规单层培养28.9.2 克隆培养28.10 传代28.10.1 收获率低或生长缓慢28.10.2 生长不均匀28.11 克隆28.11.1 培养皿形成的克隆数过低28.11.2 培养皿形成的克隆数太多28.11.3 非随机分布28.12 交叉污染和错误标记28.13 冻存28.13.1 复苏时存活率低28.13.2 冻存后细胞形态发生变化28.13.3 冻存细胞丢失28.14 细胞计数28.14.1 用血细胞计数板计数28.14.2 使用阻抗法电子计数仪计数第29章 结语附录附录Ⅰ 计算配制及试剂制备计算换算试剂制备附录II 术语细胞组织培养培养细胞的生物学组培实验室的设计及布局组培的设备和材料无菌技术组培实验的安全性、生物伦理及验证组织培养器皿和附着物成分明确培养基及补充物无血清培养基组培实验准备与灭菌原代培养传代培养和细胞系细胞系的身份认证和验证组培实验污染细胞系的冻存和建库克隆培养及筛选细胞分选细胞系鉴定组培中的细胞分化细胞的三维培养

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