安徽医科大学生命科学学院 School Of Life Sciences Aukui Wedieal Hniversity 细胞生物学实验之 鸡血细胞的融合 By生物学系-李超 2016-04 This document is produced by trial version of Print2Flash.Visit www.print2flash.com for more informatior
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[目的要求] 1.掌握细胞融合的概念及基本原理。 2.初步掌握PEG诱导细胞融合技术。 3.了解细胞融合技术的生物学和医学意义。 2 This document is produced by trial version of Print2Flash.Visit www.print2flash.com for more information
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[实验原理] 1.细胞融合（cell fusion)概念 指在自然条件下或用人工诱导的方法将两个 或多个细胞合并成一个细胞的过程。 ◆克隆羊多莉和它的 “父亲”维尔穆特 博士 This document is produced by trial version of Print2Flash.Visit www.print2flash.com for more information
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甲羊 胚胎移入子 宫内发育 丙羊 取出细胞核 提供乳 腺细胞 提供卵 乳腺细胞核移 细胞 入去核卵细胞发育成胚胎 去核卵细胞 虽然“多利”是丙羊生出的， 但其相貌却与丙不一样，而是与提 供细胞核的甲羊长相很相似 乙羊 web5410 This document is produced by trial version of Print2Flash.Visit www.print2flash.com for more information
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[实验原理] 2.促细胞融合的方法 生物诱导剂，如灭活的仙台病毒 细胞融合 诱导剂 化学诱导剂，如聚乙二醇(PEG) 物理因子，如电脉冲 花州U测 This document is produced by trial version of Print2Flash.Visit www.print2flash.com for more information
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[实验原理] Antigen cell-culture myeloma line Niels K.Jerne Georges J,F.K C閎ar Milstein ler fuse in polyethylene lycol☐←一聚乙二醇 propagate elones Freeze Thaw grow in mass induce tumors ◆1984年小鼠单克隆抗体制备 技术获得诺贝尔生理医学奖 ANTIBODY ANTIBODY 鱼精 This document is produced by trial version of Print2Flash.Visit www.print2flash.com for more information
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[实验原理] 3.PEG诱导细胞融合 √PEG是乙二醇的多聚化合物，存在一系列不同分子 量的多聚体。 √PEG可与水分子借氢键结合，在高浓度(50%)的PEG 溶液中自由水消失，导致细胞脱水而发生质膜结 构的变化，引起细胞融合。 √ 为了发挥PEG促进细胞融合的效力，必须采用较高 浓度的PEG溶液，但在高浓度PEG溶液下，细胞可 能因脱水而受到显著的破坏。因此，选择合适的 分子量、浓度及作用时间是PEG融合技术的关键。 This document is produced by trial version of Print2Flash.Visit www.print2flash.com for more information
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本期实验内容 鸡血细胞的融合 ◆ 实验材料特点：鸡血 细胞核结构致密，易于 观察。 This document is produced by trial version of Print2Flash.Visit www.print2flash.com for more information
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[实验用品] 材料：新鲜鸡血与Alsever液的混合液 器具：普通光学显微镜、离心机、恒温水浴箱、5ml 刻度离心管、试管架、一次性吸管、载玻片、盖玻片等。 试剂：50%PEG(现用现配，使用PEG4000和GKN溶 液配制)，Alsever溶液，0.75%生理盐水，GKN溶液， 0.2%的詹那斯绿B。 This document is produced by trial version of Print2Flash.Visit www.print2flash.com for more information
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[实验步骤] 1.离心洗涤 取(1：4)鸡血悬液1ml 加0.75%生理盐水至4ml,混匀 0 重复1次 3000rpm, 离心1min,弃上清 0 加GKN溶液至4l,混匀 3000rpm, 离心1min,弃上清 This document is produced by trial version of Print2Flash.Visit www.print2flash.com for more information
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