PEG介导细胞融合最适条件探讨
Optional Conditions of PEG-Induced Cell Fusion
Abstract: Cell fusion is a basic content and cell engineering technology, including PEG (polyethylene glycol) is a kind of chemical inducers, which is widely used in the modern teaching and research. This article mainly took chicken blood red blood cells for research materials, and PEG as inducers. In order to study the influence of PEG treatment on chicken red blood cell fusion, different PEG concentrations and different PEG-induced time was set in this experiment. The experimental results showed that the increase in chicken blood erythrocyte aggregation rate was observed with the increase in PEG treatment time, and that the increase in chicken blood red blood cell fusion rate was detected with increased concentrations of PEG, the concentration of which ranged from 30% to 50%. The best concentration of PEG (50%) facilitated the chicken blood red blood cell fusion. However, when the concentration was beyond 50%, chicken blood erythrocyte aggregation rate declined. In conclusion, the best concentration of PEG is 50%, and the best treatment time is 20 min for the chicken blood red blood cell fusion.
Key words: chicken blood red blood cells; Cell fusion; PEG; Fusion rate
目 录
摘 要 1
引言 2
1 实验仪器､材料和试剂 2
1.1仪器 2
1.2材料 2
1.3试剂 2
1.3.1 0.85%生理盐水 2
1.3.2 GKN液 2
1.3.3 Hank原液(10×) 2
1.3.4 Hank液 3
1.3.5不同浓度的PEG溶液(m/V) 3
1.3.6 1%Janus green B染液 3
2 方法与步骤 3
2.1鸡血细胞悬浮液的制备 3
2.2步骤 4
2.2.1不同PEG浓度对细胞融合的影响 4
2.2.2不同PEG处理时间对细胞融合的影响 4
2.3融合率的计算 5
3 结果与分析 5
3.1 PEG浓度对细胞融合的影响 5
3.2 不同PEG处理时间对细胞融合的影响 5
4 讨论 6
参考文献 7
致 谢 8
PEG介导细胞融合最适条件探讨
引言
细胞融合(cell fusion)又称体细胞杂交,指通过化学､物理等方法将两个或两个以上的体细胞融合成一个杂种细胞(hybrid cell)的现象[ ]｡细胞融合技术在细胞工程方面有重要应用,如在遗传､基因定位､细胞免疫方面是一项基础技术,同时细胞融合也是病毒､肿瘤以及培育生物新品种的重要基础手段,对其后续工作中起着奠基性作用,因而具有重要的实践意义[ ]｡可以说细胞融合技术是从事细胞方面研究必须掌握的一项基本技能｡
常规的细胞融合的方法主要有仙台病毒诱导法､电融合法(物理法)､PEG(聚乙二醇)法等[ ]｡其中由于PEG法诱导细胞融合对仪器设备的要求比较低､实验步骤操作简单､方便,血红细胞的活性比较稳定,相对其他方法比较容易制备｡PEG诱导法在生物､基因工程､医药等科研领域得到广泛应用[ ]｡Mg2+浓度､Ca2+浓度､细胞融合时的温度､PEG处理时间､PEG分子大小等诸多因素会影响细胞融合的效果[ ] [ ] [ ]｡所以,研究PEG介导细胞融合的最适条件对细胞学实验教程的优化,以及科学研究有实际应用价值[ ]｡本文以鸡血红细胞为实验材料,从不同PEG浓度和不同PEG处理时间两个方面,来探讨PEG介导细胞融合的最适条件｡从而为细胞融合实验教程和相关细胞融合的科学研究提供一定的参考｡