[Short term stimulation of megakaryopoiesis in cord blood derived hematopoietic stem cells in ex vivo model].
Ex vivo expansion of megakaryocytes may be useful to prevent the long-term severe thrombocytopenia following umbilical cord blood transplantation. This study aimed to establish the optimal conditions of short-term stimulation of megakaryopoiesis. Immunomagnetically isolated CD34 cells of umbilical cord were placed into serum free culture medium supplemented with combinations of growth factors as follow: Tpo (thrombopoietin) and IL-3 (interleukin 3); Tpo, IL-3 and SCF (stem cell factor) or Tpo, II-3, SCF and IL-11 (interleukin 11). The number of colonies of megakaryocytic cells (expressed CD41 antigen) were evaluated in clonal culture in 24-hours intervals up to 5 days. Besides the influence of used culture systems on the other hematopoietic lines was checked on the same time points. It was shown that the best results were obtained using combination of 4 cytokines. At 5-th day of expansion the number of CD41 positive cells increased in this warrant 8-fold. In case of erythroid or granulocyte-macrophage precursors the use of this combination of growth factors resulted in slightly elevated number of them. We can conclude that proposed model of expansion of megakaryopoiesis seems to be very effective and can to be useful for the shortening the post transplant thrombocytopenia period.
Machaj
Eugeniusz K
EK
Zakład Hematologii Doświadczalnej, Centrum Onkologii, Instytut im. M. Skłodowskiej-Curie, Zakład Hematologii Radiacyjnej, WIHiE w Warszawie. ekmachaj@coi.waw.pl
Gajkowska
Agnieszka
A
Jastrzewska
Marzena
M
Ołdak
Tomasz
T
Kruszewski
Marcin
M
Pojda
Zygmunt
Z
D018952:Antigens, CD34 / Q000378:metabolism*; D002452:Cell Count; D002455:Cell Division; D002469:Cell Separation / Q000379:methods; D036101:Cord Blood Stem Cell Transplantation / Q000009:adverse effects; D016895:Culture Media, Serum-Free; D005312:Fetal Blood / Q000378:metabolism*; D006801:Humans; D017370:Interleukin-11 / Q000378:metabolism; D007377:Interleukin-3 / Q000378:metabolism; D008280:Magnetics; D008533:Megakaryocytes / Q000276:immunology / Q000378:metabolism*; D019089:Stem Cell Factor / Q000378:metabolism; D013921:Thrombocytopenia / Q000209:etiology / Q000517:prevention & control*; D034061:Thrombopoiesis / Q000276:immunology; D013926:Thrombopoietin / Q000378:metabolism; D013997:Time Factors
D018952:Antigens, CD34; D016895:Culture Media, Serum-Free; D017370:Interleukin-11; D007377:Interleukin-3; D019089:Stem Cell Factor; D013926:Thrombopoietin