Flow cytometric analysis of hydroxyurea effects on fetal hemoglobin production in cultures of beta-thalassemia erythroid precursors

Hemoglobin
Ilana AmoyalEitan Fibach

Abstract

An increase in fetal hemoglobin (Hb F) ameliorates the clinical symptoms of the underlying disease in the beta hemoglobinopathies-sickle cell anemia and beta-thalassemia (thal). Hydroxyurea (HU) can elevate Hb F production in erythroid cells and is the agent currently in clinical use for patients with sickle cell anemia; it is presently being tested in clinical trials for thalassemia. We have developed a two-phase liquid culture system that mimics the in vivo hematological changes that are observed in patients treated with HU. Adding HU during the second phase of the culture increases the proportion of Hb F, increases the levels of total hemoglobin (Hb) content per cell and increases cell size, but it decreases the numbers of cells and the total amount of Hb produced. In the present study we developed and utilized a double labeling procedure for flow cytometric analysis of the cellular distribution of Hb F. Cells exposed to various concentrations of HU on day 6 of the second phase of the culture were harvested on day 12, and stained simultaneously with fluorochrome-conjugated monoclonal antibodies specific for human glycophorin A, an erythroid specific marker, and human Hb F. Both the percentage of the Hb F-containing cells and...Continue Reading

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Citations

Mar 9, 2007·Neonatology·Ilana Amoyal, Eitan Fibach
May 28, 2004·Clinical and Laboratory Haematology·I Amoyal, E Fibach
Jan 9, 2008·American Journal of Hematology·Ariel KorenHannah Tamary
Sep 30, 2008·American Journal of Hematology·Sylvia T SingerUNKNOWN E/beta Thalassemia Study Group
Jun 29, 2012·Transfusion·Eugenia Prus, Eitan Fibach
Nov 7, 2007·Journal of Pediatric Hematology/oncology·Antonis Kattamis
Apr 25, 2008·Current Protocols in Immunology·E Fibach, E Prus
Mar 1, 2008·Personalized Medicine·Marina Kleanthous, Marios Phylactides

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