Flow cytometric-based isolation of nucleated erythroid cells during maturation: an approach to cell surface antigen studies.
Cytometry
; 50(6): 305-12, 2002 Dec 15.
Article
em En
| MEDLINE
| ID: mdl-12497592
ABSTRACT
Nucleated red blood cells (NRBCs) are involved in normal physiologic processes, as well as in several malignancies. They are usually counted manually under the microscope. However, blood sample manipulation may be a source of variability and manual counting is imprecise, time-consuming, and subjective. To improve identification of CD45-negative cells, we used a flow cytometry technique that avoids the addition of lysing reagents and stains viable cell nuclei. We applied this method for counting and isolating NRBC subpopulations in whole blood samples, using DNA/RNA viable staining to discriminate nonnucleated erythroid cells and debris. NRBC counts gave 197.95 cells per mm(3) in mobilized peripheral blood samples (1.00%, n = 20), 3897.59 cells per mm(3) in leukapheresis products (3.08%, n = 20), and 765.21 cells per mm(3) in cord blood samples (6.09%, n = 20). Normal bone marrow counts were 5449.42 cells per mm(3) (11.76%, n = 20). Scatter profiles showed three distinct populations, from early to late-stage erythroblasts, consisting of erythroblasts, orthochromatic erythroblasts, and ejected nuclei, as confirmed by Wright-Giemsa staining. In addition, flow cytometry immunophenotyping showed that glycophorin A was expressed dimly on NRBCs during maturation. These findings point to the feasibility of live NRBCs studies, which offer great potential for a wide range of disciplines.
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Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Eritroblastos
/
Citometria de Fluxo
/
Antígenos de Superfície
Tipo de estudo:
Prognostic_studies
Limite:
Humans
Idioma:
En
Revista:
Cytometry
Ano de publicação:
2002
Tipo de documento:
Article
País de afiliação:
Espanha