Comparison of four serum-free, cytokine-free media for analysis of endogenous erythroid colony growth in polycythemia vera and essential thrombocythemia

The Hematology Journal : the Official Journal of the European Haematology Association
I DoboS Hermouet

Abstract

The assay of endogenous erythroid colony formation (EEC), a characteristic of polycythemia vera and essential thrombocythemia, is not standardized. In this multicentric study, we tested four semisolid, serum-free, cytokine-free media based on either methylcellulose (M1, M2) or collagen (C1, C2) commercialized for the EEC assay. Bone marrow mononuclear cells (BMMC) from 73 individuals (62 patients with either polycythemia vera (26), essential thrombocythemia (19), secondary polyglobuly (17) or chronic myeloid leukemia (2) and 11 healthy donors) were grown in parallel in the four media without, or with 0.01 U/ml erythropoietin (EPo). In all four media EEC formation was specific, as it was not observed in cultures of patients with secondary polyglobuly or chronic myeloid leukemia, nor of healthy donors. Analysis of fresh or MGG-stained collagen gel cultures allowed detection of EEC formation significantly more frequently than methylcellulose-based media; addition of 0.01 U/ml of EPo had little or no effect on EEC formation. Collagen-based medium C1 gave better results than the other media tested: the 'C1' EEC assay was positive for 68.2% of polycythemia vera cultures with significantly higher median EEC numbers (6.5/10(5) BMMC for...Continue Reading

Citations

Dec 21, 2002·British Journal of Haematology·François GirodonUNKNOWN Groupe de Standardization des Techniques de Culture de Progéniteurs Hématopoïétiques
Apr 2, 2005·British Journal of Haematology·Philipp S GoerttlerHeike L Pahl
Sep 1, 2007·Expert Opinion on Medical Diagnostics·Brijesh Arora, Ayalew Tefferi
Apr 2, 2005·Annals of Hematology·Juergen ThieleVolker Diehl
Aug 2, 2006·Archives of Pathology & Laboratory Medicine·Ming CaoYouli Zu
Feb 14, 2003·Mayo Clinic Proceedings·Ayalew Tefferi

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