Lymphocyte immunophenotyping by flow cytometry in normal adults. Comparison of fresh whole blood lysis technique, Ficoll-Paque separation and cryopreservation.
In the present report we have assessed the extent to which Ficoll-Paque separation and cryopreservation of mononuclear cells alter the measurement of lymphocyte subsets by flow cytometry. Standard Ficoll-Paque separation increased the percentage of CD4+, CD19+ and CD4+CD45RA+ cells, as well as decreasing that of CD8+, and CD4+CD29+ cells, compared to the fresh whole blood lysis technique. Moreover, cryopreservation caused a depletion of CD4+ p80+ cells, but normal whole blood values were restored following a short incubation.
Romeu
M A
MA
Servei d'Immunologia, Hospital Duran i Reynals, Barcelona, Catalunya, Spain.
Mestre
M
M
González
L
L
Valls
A
A
Verdaguer
J
J
Corominas
M
M
Bas
J
J
Massip
E
E
Buendia
E
E
D000328:Adult; D015703:Antigens, CD / Q000032:analysis; D015925:Cryopreservation; D005260:Female; D005362:Ficoll / Q000009:adverse effects; D005434:Flow Cytometry / Q000379:methods*; D006801:Humans; D016130:Immunophenotyping / Q000379:methods*; D008214:Lymphocytes / Q000276:immunology*; D008297:Male; D012680:Sensitivity and Specificity
D015703:Antigens, CD; D005362:Ficoll