Abstract: In order to solve the problem of the low effect of polyethylene glycol（PEG）-induced cell fusion, chicken blood cells were selected as experimental materials, and the fusion rate of blood cells was improved by optimizing the conditions of PEG-induced cell fusion, that is, at the fusion temperature of 38 ℃ and fusion time of 10 min, the effects of PEG relative molecular mass（2 000, 4 000, 6 000） and concentration（25%, 50%, 65%, 80%） and their interaction on hemocyte fusion rate were studied by two-factor design.The effects of fusion temperature（35, 38, 41 ℃） and time（5,10,20 min） on hemocyte fusion rate were studied by the same experimental design after determining the optimal relative molecular mass and concentration of PEG.The results showed that the relative molecular mass of PEG（F=5.36, P=0.009 2） and the concentration of PEG（F=28.46, P＜0.000 1） had a significant effect on the hemocyte fusion rate. The fusion rate of PEG-induced blood cells with relative molecular mass of 4 000 and concentration of 50% is the highest.The interaction effect of PEG relative molecular mass and concentration on the hemocytefusion rate（F=2.42, P=0.054 7）.The fusion temperature has a significant effect on the hemocyte fusion rate（F=6.56, P=0.004 8）, and the cell fusion rate was the highest at 38 ℃. However, the fusion time（F=0.34, P=0.720 0） and the interaction effect with temperature（F=0.58, P=0.680 0） had no significant effect on the hemocyte fusion rate. The results indicated that PEG relative molecular mass, concentration and fusion temperature were the key factors affecting the fusion rate of red blood cells.