A combination of megakaryocyte growth and development factor and interleukin-1 is sufficient to culture large numbers of megakaryocytic progenitors and megakaryocytes for transfusion purposes.
Chemotherapy-induced thrombocytopenia is a major risk factor in cancer treatment. The transfusion of autologous ex vivo expanded megakaryocytes could be a new therapy to shorten the period of thrombocytopenia. Therefore we investigated, in a liquid culture system, the effect of various cytokine combinations composed of pegylated megakaryocyte growth and development factor (PEG-rHuMGDF), interleukin-1 (IL-1), IL-3, IL-6, IL-11 and stem cell factor (SCF) on the proliferation and differentiation of CD34+ cells, in order to define the most optimal and minimum levels of cytokine combinations for megakaryocyte expansion. Besides PEG-rHuMGDF, IL-1 was found to be important for optimal megakaryocyte expansion. Depletion of either SCF, IL-6 or IL-11 did not exert a large effect, but the absence of IL-1 strongly diminished the number of megakaryocytic cells. Addition of IL-3 to the combination PEG-rHuMGDF, IL-1, IL-6, IL-11 and SCF significantly reduced the number of megakaryocyte progenitors (CD34+CD41+ cells) and the number of CFU-Meg. Furthermore, we found a strong correlation between the number of CD34+CD41+ cells and the number of CFU-Meg obtained after 8 d culture. Our study shows that optimal ex vivo expansion of megakaryocytes is achieved by the combination of PEG-rHuMGDF and IL-1. The numbers of megakaryocytes and megakaryocyte progenitors (CD34+CD41+) obtained in our liquid culture system with the growth factor combination PEG-rHuMGDF and IL-1 are suitable for transfusion purposes.
van den Oudenrijn
S
S
Department of Experimental Immunohaematology, CLB and Laboratory of Experimental and Clinical Immunology, Academic Medical Centre, University of Amsterdam, Amsterdam, The Netherlands.
de Haas
M
M
Calafat
J
J
van der Schoot
C E
CE
von dem Borne
A E
AE
D018952:Antigens, CD34 / Q000032:analysis; D002452:Cell Count; D002455:Cell Division; D002478:Cells, Cultured; D004338:Drug Combinations; D005434:Flow Cytometry; D019650:Hematopoietic Stem Cell Mobilization / Q000379:methods*; D018380:Hematopoietic Stem Cell Transplantation / Q000379:methods; D006801:Humans; D007375:Interleukin-1 / Q000627:therapeutic use*; D008533:Megakaryocytes / Q000166:cytology*; D016253:Microscopy, Immunoelectron; D010641:Phenotype; D019039:Platelet Glycoprotein GPIIb-IIIa Complex / Q000032:analysis; D011003:Ploidies; D013926:Thrombopoietin / Q000627:therapeutic use*
D018952:Antigens, CD34; D004338:Drug Combinations; D007375:Interleukin-1; D019039:Platelet Glycoprotein GPIIb-IIIa Complex; D013926:Thrombopoietin