细胞培养技术制药资料.ppt

1 、动物细胞培养定义 动物细胞与组织培养是从动物体内取出细胞或者组织，模拟体内的生理环境，在无菌、适温和丰富的营养条件下，使离体细胞或者组织生存、生长并维持结构和功能的一门技术。 是动物细胞工程的基础。 细胞培养技术在遗传学、免疫学、肿瘤学、病毒学、分子生物学等领域已得到广泛的应用。 2、 动物细胞特点 无细胞壁 倍增时间长，生长缓慢 需氧量少，对搅拌敏感 聚集体形成 原代细胞培养50代即开始退化 3、 发展历史 1907，美国的哈里森使用盖玻片悬滴培养蛙胚神经组织细胞 4、 生长特性 贴附生长型 如人胚肺细胞，Hela细胞，巨噬细胞，神经细胞 悬浮生长型细胞 如血液白细胞，淋巴组织细胞等 5、 体外培养细胞基本技术 体外培养特点 体外培养工具 体外培养条件 体外细胞生长增殖过程 培养方法 大规模培养技术 5.1 体外培养特点 营养条件苛刻 适应性差, 敏感 培养时间长，易污染 Animal cells are more difficult to culture than microorganisms because they require many more nutrients and typically grow only when attached to specially coated surfaces. Despite these difficulties, various types of animal cells, including both undifferentiated and differentiated ones, can be cultured successfully. 5.3 体外培养条件 温度，37度 pH：7.2-7.4 气体，氧，二氧化碳，氮气 营养条件 需要多种氨基酸，维生素，辅酶，核酸，嘌呤，嘧啶，激素和生长因子，其中多种成分可以由血清提供 培养条件 无菌、无毒 消毒、灭菌 抗生素 定期更换培养液，清除代谢废物 营养 合成培养基：糖、氨基酸、生长因子、无机盐、微量元素 （血清、血浆） 温度和pH 36.5±0.5℃，7.2---7.4 气体 95%空气，5%CO2 Rich Media Are Required for Culture of Animal Cells Nine amino acids, referred to as the essential amino acids, cannot be synthesized by adult vertebrate animals and thus must be obtained from their diet. Animal cells grown in culture also must be supplied with these nine amino acids, namely, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, threonine, tryptophan, and valine. addition, most cultured cells require cysteine, glutamine, and tyrosine. The other essential components of a medium for culturing animal cells are vitamins, which the cells cannot make at all or in adequate amounts; various salts; glucose; and serum, the noncellular part of the blood Most Cultured Animal Cells Grow Only on Special Solid Surfaces The extracellular matrices in various animal tissues consist of several common components: fibrous collagen proteins; hyaluronan (or hyaluronic acid), a large mucopolysaccharide; and covalently linked polysaccharides and proteins in the form of proteoglycan (mostly carbohydrat