PURIFICATION OF HEMATOPOIETIC PROGENITOR CELLS FROM HUMAN MARROW USING A FUCOSE-BINDING LECTIN AND CELL SORTING

Abstract

Human peripheral blood granulocytes, but not lymphocytes, erythrocytes or monocytes, bound the fucose-binding lectin from Lotus tetragonolobus (FBP), and this binding was competitively inhibited by the sugar .alpha.-L-fucose. The fluorescence-activated cell sorter was used to study the appearance of this receptor on human marrow cells during granulocyte differentiation and to prepare fractions enriched for granulocyte-macrophage progenitor cells (granulocyte-macrophage colony-forming cells-GM-CFC). Cell binding of fluoresceinated FBP increased for bone marrow cells in the sequence.sbd.lymphocytes, blast cells, promyelocytes and myelocytes, monocytes and polymorphonuclear cells. Selection of cells with appropriate low-angle or high-angle light scatter characteristics achieved a 10-fold or 2- to 3-fold enrichment of progenitor cells, respectively. By selecting cells with intermediate fluorescence intensity, a further 2- to 3-fold enrichment for GM-CFC was obtained. Cell sorting using the optimal selection of these 3 parameters produced up to 36-fold enrichment of the progenitor cells from human bone marrow. The most enriched fraction was composed of 23% progenitor cells (colony- and cluster-forming cells) with a yield of 36%. In populations most highly enriched for GM-CFC, immature cells (blast cells, promyelocytes and myelocytes) made up 95% of the cells present.