2009International Journal of Laboratory HematologyRequires access

A biparametric flow cytometry analysis for the study of reticulocyte patterns of maturation

Paolo Borrione, Angela Spaccamiglio, Attilio Parisi, R. A. Salvo, Marisa Pautasso, Fabio Pigozzi, Alberto Angeli

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Abstract

Automated haematological analysers still represent the gold standard for the study of reticulocyte maturation even if this technique is based on structural properties and staining affinity rather than on functional aspects. On the contrary, flow cytometry allows the simultaneous analysis of multiple cellular characteristics including functional features. Aim was to investigate whether simultaneous analysis of different reticulocyte parameters using flow cytometry may add functional information when considering their pattern of maturation. Thirty-nine healthy donors (H) and 31 haemodialysed patients on treatment with rHuEpo (HDT) were analysed. Reticulocyte counts and their stages of maturation were studied both with ADVIA 2120 and by flow cytometry. TO/CD71 scattergraph reticulocyte analysis designed a peculiar distribution which was similar among the same group of subjects (H or HDT), but different between H and HDT. distribution of the percentage of reticulocytes in low, medium and high boxes calculated by ADVIA 2120 did not show any difference between H and HDT groups, while the analysis using flow cytometry pointed out statistically significant differences between H and HDT groups in the three boxes where the TO+/CD71+ reticulocytes were localized. The present study suggests that TO/CD71 analysis was reproducible and could detect different pattern of maturation of a particular clinical setting.

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What this paper is about

Automated haematological analysers still represent the gold standard for the study of reticulocyte maturation even if this technique is based on structural properties and staining affinity rather than on functional aspects. On the contrary, flow cytometry allows the simultaneous analysis of multiple cellular characteristics including functional features. Aim was to investigate whether simultaneous analysis of different reticulocyte parameters using flow cytometry may add functional information when considering their pattern of maturation. Thirty-nine healthy donors (H) and 31 haemodialysed patients on treatment with rHuEpo (HDT) were analysed. Reticulocyte counts and their stages of maturation were studied both with ADVIA 2120 and by flow cytometry. TO/CD71 scattergraph reticulocyte analysis designed a peculiar distribution which was similar among the same group of subjects (H or HDT), but different between H and HDT. distribution of the percentage of reticulocytes in low, medium and high boxes calculated by ADVIA 2120 did not show any difference between H and HDT groups, while the analysis using flow cytometry pointed out statistically significant differences between H and HDT groups in the three boxes where the TO+/CD71+ reticulocytes were localized. The present study suggests that TO/CD71 analysis was reproducible and could detect different pattern of maturation of a particular clinical setting.

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Available abstract

Automated haematological analysers still represent the gold standard for the study of reticulocyte maturation even if this technique is based on structural properties and staining affinity rather than on functional aspects. On the contrary, flow cytometry allows the simultaneous analysis of multiple cellular characteristics including functional features. Aim was to investigate whether simultaneous analysis of different reticulocyte parameters using flow cytometry may add functional information when considering their pattern of maturation. Thirty-nine healthy donors (H) and 31 haemodialysed patients on treatment with rHuEpo (HDT) were analysed. Reticulocyte counts and their stages of maturation were studied both with ADVIA 2120 and by flow cytometry. TO/CD71 scattergraph reticulocyte analysis designed a peculiar distribution which was similar among the same group of subjects (H or HDT), but different between H and HDT. distribution of the percentage of reticulocytes in low, medium and high boxes calculated by ADVIA 2120 did not show any difference between H and HDT groups, while the analysis using flow cytometry pointed out statistically significant differences between H and HDT groups in the three boxes where the TO+/CD71+ reticulocytes were localized. The present study suggests that TO/CD71 analysis was reproducible and could detect different pattern of maturation of a particular clinical setting.

Key concepts: Reticulocyte, Flow cytometry, Cytometry, Biology, Andrology, Medicine, Molecular biology, Biochemistry

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