Multiparametric analysis for the enumeration of CD34+ cells from bone marrow and stimulated peripheral blood.
Determination of CD34+ cells was performed in bone marrow and G-CSF mobilised peripheral blood samples. We adopted three different protocols of analysis: the Milan/Mulhouse protocol, the ISHAGE guidelines for CD34+ cell determination and our own protocol based upon the use of PAINT-A-GATEPRO software analysis program. An excellent correlation was demonstrated between the three methods (r2 0.98); however the analysis of variance showed a statistically significant difference between the results generated with the three methods (P=0.001). The differences between the three procedures are discussed with a special focus on the value of CD34+dim cells and the role of CD45 in the setting of a double staining. We have in fact identified a minor subset (CD34+CD38+CD45-) which would go unrecognised based upon its CD45 negativity.
Venditti
A
A
Cattedra di Ematologia, Universita 'Tor Vergata', Divisione di Ematologia, Ospedale S. Eugenio, Piazzale Umanesimo 10, 00144 Rome, Italy.
Buccisano
F
F
Del Poeta
G
G
Battaglia
A
A
Tamburini
A
A
Bruno
A
A
Adorno
G
G
Caravita
T
T
Santinelli
S
S
Del Moro
B
B
Epiceno
A M
AM
Ballatore
G
G
Amadori
S
S
D018952:Antigens, CD34 / Q000032:analysis*; D001854:Bone Marrow Cells / Q000166:cytology* / Q000276:immunology; D002469:Cell Separation; D005434:Flow Cytometry / Q000379:methods; D006801:Humans; D007091:Image Processing, Computer-Assisted; D007963:Leukocytes, Mononuclear; D013234:Stem Cells
D018952:Antigens, CD34