安徽医科大学：《医学细胞生物学》课程教学资源（实践指导，打印版）鸡血细胞的融合

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

This document is produced by trial version of Print2Flash. Visit www.print2flash.com for more information

[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

This document is produced by trial version of Print2Flash. Visit www.print2flash.com for more information

[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

This document is produced by trial version of Print2Flash. Visit www.print2flash.com for more information

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

This document is produced by trial version of Print2Flash. Visit www.print2flash.com for more information

[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

This document is produced by trial version of Print2Flash. Visit www.print2flash.com for more information

[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

This document is produced by trial version of Print2Flash. Visit www.print2flash.com for more information

[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

This document is produced by trial version of Print2Flash. Visit www.print2flash.com for more information

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

This document is produced by trial version of Print2Flash. Visit www.print2flash.com for more information

[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

This document is produced by trial version of Print2Flash. Visit www.print2flash.com for more information

[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

This document is produced by trial version of Print2Flash. Visit www.print2flash.com for more information