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Hemopoietic stem cells (HSC) are identified through their unique ability, at the single cell level, to long-term reconstitute all blood cell lineages. Sustained myeloid reconstitution is considered the hallmark of HSC, because myeloid progenitors and their progeny have very short half-lives. Here we demonstrate that the established practice of relying on RB6-8C5 as a myeloid specific Ab can result in overestimation of HSC frequencies because the RB6-8C5 Ab also detects Ags expressed on a sizeable population of CD3(+)CD8(+) T cells, constitutively as well as following transplantation. Likewise, a high fraction of mice transplanted with limiting numbers of ex vivo expanded Lin(-)Sca(+)kit(+)CD34(-) HSC show long-term RB6-8C5(+)CD3(+) (lymphoid) but no RB6-8C5(+)CD3(-) (myeloid) reconstitution. Most noteworthy, the use of RB6-8C5 as a myeloid specific Ab can be deceptive by implicating the existence of lineage-restricted HSC capable of long-term reconstituting the myeloid and T, but not B, cell lineage. Because cross-lineage expression of "lineage-specific" markers is unlikely to be unique to the blood system, claims of unexpected cell fates should be substantiated not only by acquisition of lineage-specific markers, but also absence of markers of other lineages normally derived from the investigated stem cells.

Original publication




Journal article


J Immunol

Publication Date





1548 - 1552


Animals, Antibodies, Monoclonal, Antigens, Differentiation, B-Lymphocyte, Antigens, Differentiation, Myelomonocytic, Antigens, Differentiation, T-Lymphocyte, Blood Cell Count, CD3 Complex, CD8-Positive T-Lymphocytes, Cell Differentiation, Cell Lineage, Colony-Forming Units Assay, Epitopes, Hematopoietic Stem Cell Transplantation, Hematopoietic Stem Cells, Mice, Mice, Inbred C57BL, Myeloid Progenitor Cells, T-Lymphocyte Subsets