目的:细胞融合是细胞生物学领域近30年来得到迅速发展的一项新兴技术手段,因其操作简便、人工可控等优点在研究核质互作、肿瘤发生、疫苗研发和培育新型生物品种等方面均有广泛应用。其中,利用聚乙二醇(PEG)进行化学融合是细胞融合中最为常用且简便的技术手段。PEG化学融合效果受到多种因素影响,如PEG浓度、Ca^2+、Mg^2+、pH值等,然而对于糖类物质在细胞融合中的影响未见报道。本文旨在为了更全面了解PEG法诱导的化学细胞融合,通过优化融合条件以提高化学细胞融合效率。方法:选取鸡血血细胞为材料,通过改变原Hanks缓冲液中葡萄糖浓度,观察比较各组细胞融合率,探究葡萄糖浓度在化学细胞融合中的影响,并通过对比结果获得了对于鸡血血细胞应采用的最适葡萄糖浓度区间。结果:对于鸡血血细胞融合实验,葡萄糖浓度在10-14mmol/L范围内细胞融合效率较原Hanks液配方高2倍左右。结论:葡萄糖对细胞融合效果具有一定的影响,可以通过调节葡萄糖浓度提高细胞融合率,从而为PEG化学细胞融合提供一种更为优化的方案。
Objective: Cell fusion is one of the new cytological techniques in recent 30 years. Because it is easy to be operated and controlled, cell fusion is now widely used in many fields such as nucleoplasm research, tumor genesis, vaccine development and nurturing new biological species etc. Polyethylene glycol (PEG) chemical fusion is the most common cell fusion technology. There are many factors affecting the PEG chemical fusion such as the PEG concentration, Ca^2+, Mg^2+, pH value and et al. However, the effect of glucose concentration on cell fusion has not been reported. In order to understand the factor influencing the chemistry of PEG-induced cell fusion better, we optimized fusion conditions, and improved the fusion rate of the PEG chemical fusion. Methods: Choose chicken red blood cells as material, and vary the concentrations of glucose in the original Hanks buffer. By comparing with the cell fusion rate in different glucose concentration, the effect of glucose concentration on the cell fusion and the better glucose concentration for fusion rate were explored. Results: Based on the original Hanks buffer recipe in chicken cell fusion, we adjusted the concentration range of glucose to 10 mmol/L-14 mrnol/L, which contributed to a higher rate of cell fusion. Conclusion: Glucose concentration has an effect on cell fusion and higher concentration of glucose is better for cell fusion. This work provides an optimization for the PEG chemical fusion.