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      Isolation of single human hematopoietic stem cells capable of long-term multilineage engraftment.

      Science (New York, N.Y.)
      Animals, Antigens, CD34, analysis, Antigens, Thy-1, Cell Lineage, Cell Proliferation, Cell Separation, Coculture Techniques, Fetal Blood, cytology, Flow Cytometry, Gene Expression Profiling, Hematopoiesis, Hematopoietic Stem Cell Transplantation, Hematopoietic Stem Cells, immunology, physiology, Humans, Integrin alpha6, Mice, Mice, Inbred NOD, Multipotent Stem Cells, Stromal Cells, Transplantation, Heterologous

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          Abstract

          Lifelong blood cell production is dependent on rare hematopoietic stem cells (HSCs) to perpetually replenish mature cells via a series of lineage-restricted intermediates. Investigating the molecular state of HSCs is contingent on the ability to purify HSCs away from transiently engrafting cells. We demonstrated that human HSCs remain infrequent, using current purification strategies based on Thy1 (CD90) expression. By tracking the expression of several adhesion molecules in HSC-enriched subsets, we revealed CD49f as a specific HSC marker. Single CD49f(+) cells were highly efficient in generating long-term multilineage grafts, and the loss of CD49f expression identified transiently engrafting multipotent progenitors (MPPs). The demarcation of human HSCs and MPPs will enable the investigation of the molecular determinants of HSCs, with a goal of developing stem cell-based therapeutics.

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