Purification of rabbit blood basophils by isopycnic banding on Percoll.
This study defines the basophil densities in rabbits and also describes a relatively simple method of purifying basophils from peripheral blood through a two-step procedure. To evaluate the recovery of basophils, EDTA-anticoagulated blood in layered over Percoll of various densities, ranging from 1.070 to 1.080 g/ml. The cells retained in Percoll were collected separately after centrifugation at 600 g(av) for 30 min (Step I). Distribution of basophils indicated that basophils had densities of between 1.070 and 1.078 g/ml, with a peak at 1.074 g/ml. To enhance the basophil purity, a second centrifugation of the cells obtained from Step I over appropriate Percoll gradient led to further enhancement of basophil purity (30.4 +/- 0.7%) which was (20.6 +/- 0.2%) in Step I. The cells obtained by this method appeared morphologically normal and viable.
Department of Immunology, ICMR, Kobe University School of Medicine, Japan.
M
M
Nouri-Shirazi
K
K
Fukuta
Y
Y
Ohta
D000818:Animals; D001491:Basophils; D002469:Cell Separation / Q000379:methods*; D002500:Centrifugation, Isopycnic / Q000379:methods; D003102:Colloids; D007958:Leukocyte Count; D011205:Povidone; D011817:Rabbits; D012822:Silicon Dioxide
D003102:Colloids; C016039:Percoll; D012822:Silicon Dioxide; D011205:Povidone