Superior leukocyte separation with a discontinuous one-step Ficoll-Hypaque gradient for the isolation of human neutrophils.
A discontinuous gradient system composed of two commercially available Ficoll-Hypaque mixtures (Mono-Poly resolving medium (MPRM) and Histopaque 1.077) is described for the purification of mononuclear and polymorphonuclear leukocytes from human blood. Like the original one-step Hypaque-Ficoll procedure (Ferrante and Thong, 1978), a single centrifugation at 500 X g for 30 min resulted in the formation of two distinct leukocyte fractions. In contrast to MPRM alone, the discontinuous system (MPRM-HP) was capable of resolving leukocyte fractions from blood volumes as small as 1 ml with excellent purity and yield. MPRM-HP was also compatible with a wider range of anticoagulants and permitted fractionation of specimens resistant to MPRM.
Kalmar
J R
JR
Department of Oral Biology, Emory University Dental Research Center, Atlanta, GA 30322.
Arnold
R R
RR
Warbington
M L
ML
Gardner
M K
MK
D000328:Adult; D002469:Cell Separation / Q000379:methods*; D002499:Centrifugation, Density Gradient / Q000379:methods*; D019343:Citric Acid; D003973:Diatrizoate; D005260:Female; D005362:Ficoll; D005947:Glucose / Q000031:analogs & derivatives; D006493:Heparin; D006801:Humans; D007202:Indicators and Reagents; D009504:Neutrophils; D009994:Osmolar Concentration; D011134:Polysaccharides
D007202:Indicators and Reagents; D011134:Polysaccharides; D003973:Diatrizoate; C002113:acid citrate dextrose; D005362:Ficoll; D019343:Citric Acid; D006493:Heparin; D005947:Glucose