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Effects of recombinant thrombopoietin on the growth of murine primitive and committed hematopoietic progenitors in serum‐free culture
Author(s) -
Sasaki HIDEKI,
Ikuta KOICHIRO,
Funabiki TETSUNORI,
Fujioka KENICHIRO,
Matsuda MOTOI
Publication year - 1999
Publication title -
pediatrics international
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.49
H-Index - 63
eISSN - 1442-200X
pISSN - 1328-8067
DOI - 10.1046/j.1442-200x.1999.01141.x
Subject(s) - thrombopoietin , haematopoiesis , megakaryocyte , progenitor cell , clonogenic assay , erythropoietin , recombinant dna , interleukin 3 , granulocyte macrophage colony stimulating factor , microbiology and biotechnology , colony stimulating factor , immunology , stem cell factor , macrophage , granulocyte , biology , cell culture , stem cell , cytokine , endocrinology , in vitro , biochemistry , t cell , genetics , immune system , antigen presenting cell , gene
Background : The aim of the present study was to determine the effect of thrombopoietin (Tpo) in combination with other cytokines on the growth of murine megakaryocytic, granulocyte–macrophage, erythroid and primitive hematopoietic progenitor cells, excluding possibilities of synergistic effects by serum factor(s). Methods : Serum‐free clonogenic assay systems were used for assay of colony forming units in megakaryocytes (CFU‐Mk), colony forming units in granulocytes‐macrophages (CFU‐GM), burst‐forming units in erythrocytes (BFU‐E) in marrow of normal mice and of high proliferative potential colony forming cells in 5‐fluorouracil‐treated marrow. Results : Serum‐free culture enabled megakaryocyte colony growth by recombinant murine (rm) Tpo and this was synergistically supported with rm interleukin (IL)‐3, rm stem cell factor (SCF) or recombinant human (rh) erythropoietin (Epo). Recombinant human IL‐6, rhIL‐11 and rm granulocyte–macrophage colony stimulating factor (GM‐CSF) showed synergistic effects with rmTpo only in the presence of serum. Recombinant murine IL‐3 or rmSCF increased the large colonies and mixed‐type colonies containing other populations, suggesting that these cytokines promoted the proliferation of immature populations of CFU‐Mk. The rmTpo enhanced the growth of granulocyte–macrophage (GM) colonies stimulated by rmGM‐CSF or rmIL‐3 and erythroid bursts by rhEpo, with or without rmIL‐3. The rmTpo also significantly increased HPP colonies in synergy with rmIL‐3 only in the presence of serum or rmSCF. Conclusion : Serum‐free culture is valuable for evaluating synergistic effects of cytokines and Tpo acts not only on megakaryocytic progenitors but also on granulocyte‐macrophage, erythroid and primitive progenitor cells in combination with other cytokines, such as rmIL‐3 and rmSCF. However, serum or SCF was required for enhancement of the colony growth of primitive progenitors by Tpo.