[Method for human peripheral blood eosinophil isolation for patch-clamp study].
OBJECTIVE
To establish a rapid and economic method for isolating human peripheral blood eosinophils with high viability for patch-clamp studies and investigate the electrophysiological properties of Ca(2+)-activated K(+) channel of the isolated cells.


METHODS
Peripheral blood eosinophils were isolated by modified discontinuous Percoll density gradient centrifugation, and the electric currents in the single Ca(2+)-activated K(+) channels of the cells were recorded using patch-clamp technique with cell-attached configuration.


RESULTS
The purity of the eosinophils from healthy donors reached (90.5+/-1.6)%, with a viability rate over 99% and recovery rate of (48.2+/-6.9)%. The isolated cells were morphologically intact, from which Ca(2+)-activated K(+) channel activity could be detected.


CONCLUSIONS
The peripheral blood eosinophils isolated using this rapid, simple and highly efficient method are characterized by high purity and viability without obvious cellular injuries, which are ideal for patch-clamp studies.
Zhou
Bin
B
Department of Respiratory Diseases, Nanfang Hospital, Southern Medicine University, Guangzhou 510515, China.
Cai
Shao-xi
SX
Zhao
Hai-jin
HJ
Hu
Guo-dong
GD
D002469:Cell Separation; D004804:Eosinophils / Q000166:cytology*; D006801:Humans; D018408:Patch-Clamp Techniques; D024681:Potassium Channels, Calcium-Activated / Q000032:analysis
D024681:Potassium Channels, Calcium-Activated