title: Ex vivo expansions of megakaryocytopoiesis from placental and umbilical cord blood CD34(+) cells in serum-free culture supplemented with proteoglycans extracted from the nasal cartilage of salmon heads and the nasal septum cartilage of whale.
abstract: As a possible approach to the treatment of thrombopocytopenia, the ex vivo expansion of megakaryocytic progenitor cells may be a useful tool to accelerate platelet recovery in vivo. Our objective was to assess the promoting effect of proteoglycans in a serum-free culture condition using human cord blood CD34(+) cells. Highly purified proteoglycan (PG) extracted from the nasal cartilage of salmon heads and the nasal septum cartilage of a whale were applied to the ex vivo expansion of megakaryocytopoiesis and thrombopoiesis from placental and umbilical cord blood CD34(+) cells in serum-free cultures stimulated with a combination of thrombopoietin (TPO) and interleukin-3 (IL-3). Each PG (0.5 and 5 mug) was applied to the culture with three different concentrations of TPO (50, 5 and 0.5 ng/ml) and IL-3 (100, 10 and 1 ng/ml). Both of the PGs showed no promoting effects on the mononuclear cell proliferation rate in any of the cultures. However, the whale-PG promoted the generation of megakaryocytic progenitor cells and megakaryocytes in the culture with a lower dose of cytokines, respectively. In addition, whale-PG led to a significant increase in CD42a(+) particles which seemed to be platelets. While the salmon-PG failed to promote such production in almost all of the cultures. Although whale-PG is an attractive molecule for the ex vivo expansion of human megakaryocytopoiesis, its action may depend on the glycosaminoglycans sulfation pattern and the ability of the binding affinity and the kinetics to interact with the cytokines and hematopoietic stem/progenitor cells.
authors.affiliation: Department of Radiological Life Sciences, Division of Medical Life Sciences, Hirosaki University Graduate School of Health Sciences, 66-1 Hon-cho, Hirosaki 036-8564, Japan. ikashi@cc.hirosaki-u.ac.jp
authors.forename: Ikuo
authors.initials: I
authors.lastname: Kashiwakura
authors.forename: Kenji
authors.initials: K
authors.lastname: Takahashi
authors.forename: Satoru
authors.initials: S
authors.lastname: Monzen
authors.forename: Toshiya
authors.initials: T
authors.lastname: Nakamura
authors.forename: Keiichi
authors.initials: K
authors.lastname: Takagaki
mesh_terms: D000818:Animals; D018952:Antigens, CD34 / Q000276:immunology*; D002478:Cells, Cultured; D016895:Culture Media, Serum-Free; D005260:Female; D005312:Fetal Blood / Q000166:cytology* / Q000276:immunology; D006801:Humans; D007377:Interleukin-3 / Q000494:pharmacology; D007963:Leukocytes, Mononuclear / Q000166:cytology / Q000187:drug effects* / Q000276:immunology; D008533:Megakaryocytes / Q000166:cytology / Q000187:drug effects* / Q000276:immunology; D009300:Nasal Septum / Q000737:chemistry*; D010920:Placenta / Q000098:blood supply*; D011509:Proteoglycans / Q000302:isolation & purification / Q000494:pharmacology*; D012474:Salmon; D034061:Thrombopoiesis / Q000187:drug effects* / Q000276:immunology; D013926:Thrombopoietin / Q000494:pharmacology; D014907:Whales
chemical_list: D018952:Antigens, CD34; D016895:Culture Media, Serum-Free; D007377:Interleukin-3; D011509:Proteoglycans; D013926:Thrombopoietin