Structurally Specific Heparan Sulfates Support Primitive Human Hematopoiesis by Formation of a Multimolecular Stem Cell Niche

Stem cell localization, conservation, and differentiation isbelieved to occur in niches in the marrow stromal microenvironment. Ourrecent observation that long-term in vitro human hematopoiesis requiresa stromal heparan sulfate proteoglycan (HSPG) led us to hypothesizethat such HSPG may orchestrate the formation of the stem cell niche. Wecompared the structure and function of HS from M2-10B4, ahematopoiesis-supportive cell line, with HS from a nonsupportive cellline, FHS-173-We. Long-term culture-initiating cell (LTC-IC)maintenance was enhanced by PG from supportive cells but not by PG fromnonsupportive cells (P < .005). The supportive HS weresignificantly larger and more highly sulfated than the nonsupportiveHS. Specifically, supportive HS contained higher 6-O-sulfation on theglucosamine residues. In agreement with these observations, purified6-O-sulfated heparin and highly 6-O-sulfated bovine kidney HS similarlymaintained LTC-IC. In contrast, completely desulfated heparin,N-sulfated heparin, and unmodified heparin did not support LTC-ICmaintenance. Moreover, the supportive HS promoted LTC-IC maintenancebut not differentiation of CD34⁺/HLA-DRcells into colony-forming cells (CFCs) and mature bloodcells. The supportive HS but not the nonsupportive HS bound bothcytokines and matrix components critical for hematopoiesis, includinginterleukin-3 (IL-3), macrophage inflammatory protein-1 (MIP-1),and thrombospondin (TSP). Significantly more CD34⁺ cellsadhered directly to immobilized O-sulfated heparin than to N-sulfatedor desulfated heparin. Thus, hematopoiesis-supportive stromal HSPGpossessing large, highly 6-O-sulfated HS mediate the juxtaposition ofhematopoietic progenitors with stromal cells, specific growth-promoting(IL-3) and growth-inhibitory (MIP-1 and platelet factor 4 [PF4])cytokines, and extracellular matrix (ECM) proteins such as TSP. Weconclude that the structural specificity of stromal HSPG thatdetermines the selective colocalization of cytokines and ECM componentsleads to the formation of discrete niches, thereby orchestrating thecontrolled growth and differentiation of stem cells. These findings mayhave important implications for ex vivo expansion of and gene transferinto primitive hematopoietic progenitors.