title: Formation of an adherent hematopoietic expansion culture using fucoidan.
abstract: Expansion of transplantable cord blood (CB) progenitors using a stroma requires provision of an exogenous cell source because of the low frequency of stromal precursor cells in CB. A simpler approach from a clinical regulatory perspective would be to provide synthetic extracellular matrix. The aim of this study was to characterize the effect on hematopoietic cell culture of fucoidan. The modulation of cytokine-driven hematopoietic cell expansion by fucoidan was investigated using two-level fractional and full factorial experimental designs. Mobilized peripheral blood (PB) CD34(+) cells were grown over 10 days in various combinations of FL, SCF, TPO, G-CSF, and SDF-1. Cultures were analyzed by immunophenotype. The effect of fucoidan on the divisional recruitment of CD34(+) cells was studied by CFDA-SE division tracking. Fucoidan was adsorbed by polystyrene to tissue culture plates and promoted formation of an adherent hematopoietic culture. Factorial design experiments with mobilized PB-CD34(+) cells showed that fucoidan reduced the production of CD34(+) cells and CD34(+)CXCR4(+) ratio but did not affect the production of monocytic, granulocytic, or megakaryocytic cells. The inhibitory effect of fucoidan on expansion of CB-CD34(+) cells was greater than mobilized PB. Division tracking analysis showed that CD34(+) cell generation times were lengthened by fucoidan. Fucoidan binds growth factors via their heparin-binding domain. The formation of an adherent hematopoietic culture system by fucoidan is most likely mediated by the binding of L-selectin and integrin-αMβ2 on myeloids. Fucoidan deserves further investigation as glycan scaffold that is suitable for immobilization of other matrix molecules thought to comprise blood stem cell niche.
authors.affiliation: Graduate School of Biomedical Engineering, University of New South Wales, Sydney, NSW 2052, Australia. mohammad.irhimeh@sydney.edu.au
authors.forename: Mohammad R
authors.initials: MR
authors.lastname: Irhimeh
authors.forename: J Helen
authors.initials: JH
authors.lastname: Fitton
authors.forename: Kap-Hyoun
authors.initials: KH
authors.lastname: Ko
authors.forename: Ray M
authors.initials: RM
authors.lastname: Lowenthal
authors.forename: Robert E
authors.initials: RE
authors.lastname: Nordon
mesh_terms: D018952:Antigens, CD34 / Q000378:metabolism; D000970:Antineoplastic Agents / Q000494:pharmacology; D002448:Cell Adhesion / Q000187:drug effects / Q000502:physiology; D018929:Cell Culture Techniques / Q000379:methods; D049109:Cell Proliferation / Q000187:drug effects*; D002478:Cells, Cultured; D054377:Chemokine CXCL12 / Q000494:pharmacology; D006412:Hematopoietic Stem Cells / Q000166:cytology* / Q000187:drug effects* / Q000378:metabolism / Q000502:physiology; D006801:Humans; D008533:Megakaryocytes / Q000187:drug effects / Q000378:metabolism / Q000502:physiology; D011134:Polysaccharides / Q000494:pharmacology*; D019089:Stem Cell Factor / Q000494:pharmacology
chemical_list: D018952:Antigens, CD34; D000970:Antineoplastic Agents; D054377:Chemokine CXCL12; D011134:Polysaccharides; D019089:Stem Cell Factor; C007789:fucoidan