A comparative study of eosinophil isolation by different procedures of CD16-negative depletion.
Eosinophils were isolated by the three methods of CD16-negative depletion: 1) magnetic beads, 2) fluorescence-activated cell sorter (FACS), and 3) complement reaction. Their purity, yield, and viability were compared. The second procedure produced well purity and viability (94.65 +/- 1.51% and 94.98 +/- 1.40%, respectively) but low yield of eosinophils (65.47 +/- 2.47%). The viability of cells obtained by the third procedure was not efficient (80.83 +/- 2.85%), while the purity and the yield were efficient (96.23 +/- 1.09% and 90.75 +/- 1.72%, respectively). In conclusion, the magnetic beads method (purity: 98.02 +/- 0.45%, yield: 91.05 +/- 2.43%, viability: 97.57 +/- 0.37%) was the most advantageous of these three procedures. Moreover, in the functional assay, radical oxygen products from eosinophils isolated by the procedure with complement reaction were less than with the magnetic beads or FACS procedures.
Chihara
J
J
Fourth Department of Internal Medicine, Kinki University School of Medicine, Osaka, Japan.
Kurachi
D
D
Yamamoto
T
T
Yamada
H
H
Wada
T
T
Yasukawa
A
A
Nakajima
S
S
D002469:Cell Separation / Q000379:methods*; D003165:Complement System Proteins; D004804:Eosinophils / Q000276:immunology*; D005434:Flow Cytometry; D006801:Humans; D008163:Luminescent Measurements; D008165:Luminol; D008280:Magnetics; D008863:Microspheres; D009504:Neutrophils / Q000276:immunology; D017452:Receptors, IgG / Q000032:analysis*
D017452:Receptors, IgG; D008165:Luminol; D003165:Complement System Proteins