Comparison of 4 popular methods for neutrophil isolation from human peripheral blood

Objective Percoll density gradient centrifugation,Ficoll-Hypaque density gradien centrifugation,red blood cells cracking and natural erythrocyte sedimentation method with Dextran,which are 4 frequently used methods for neutrophil separation from human peripheral blood,were compared.Methods Respectively using the 4 methods to separate neutrophil from same healthy human peripheral blood,the purity,recovery rate,and cell survival rate were compared.Results Among the 4 separation methods,the neutrophil purity from Percoll density gradient centrifugation and Ficoll-Hypaque density gradient centrifugation were more than 90%,but there was no significant difference between them(P0.05);the cell purity from red blood cells cracking and the natural erythrocyte sedimentation method with Dextran was lower than the Percoll density gradient centrifugation(P0.01),Ficoll-Hypaque density gradient centrifugation(P0.05);the neutrophil recovery rate from Percoll density gradient centrifugation,Ficoll-Hypaque density gradient centrifugation and red blood cells cracking were significantly higher than that from the natural erythrocyte sedimentation method with Dextran(P0.01,P0.01,and P0.05,respectively);Cell survival rate from Percoll density gradient centrifugation was significantly higher than Ficoll-Hypaque density gradient centrifugation(P0.05),red blood cells cracking(P0.01),and the natural erythrocyte sedimentation method with Dextran(P0.01).Conclusion Percoll density gradient centrifugation provides the most simple and efficient approach for isolation of human blood neutrophil with the high purity and viability,which is suitable to utilize in the clinical work and scientific research.