Human and rhesus macaque hematopoietic stem cells cannot be purified based only on SLAM family markers

Abstract

Various combinations of antibodies directed to cell surface markers have been used to isolate human and rhesus macaque hematopoietic stem cells (HSCs). These protocols result in poor enrichment or require multiple complex steps. Recently, a simple phenotype for HSCs based on cell surface markers from the signaling lymphocyte activation molecule (SLAM) family of receptors has been reported in the mouse. We examined the possibility of using the SLAM IntroductionThe search for hematopoietic stem cell (HSC)-specific surface markers has been a central question for functional stem cell studies and for the development of clinical applications, including transplantation and gene therapy. In mice, using a complex combination of positive and negative selection for 10 to 12 surface markers (Lin Ϫ Sca-1 ϩ c-kit ϩ Thy-1 lo ), 1 in 4.9 cells provides long-term reconstitution after intravenous injection. 1 However, the use of these complex sets of markers, alone or in combination with the established isolation scheme based on Hoechst dye efflux (side population), 2 is incompatible with in situ histologic analyses.Recently, a simple and broadly applicable method to isolate mouse HSCs has been developed based on expression of cell surface markers, which are members of the signaling lymphocyte activation molecule (SLAM) family, including CD150, CD48, and CD244. 1 In transplantation assays, 1 in 4.8 CD150 ϩ CD48 Ϫ bone marrow (BM) cells provides longterm, multilineage reconstitution in recipient mice, similar to the enrichment obtained in the complex Lin Ϫ Sca-1 ϩ c-kit ϩ Thy-1 lo population. 1 This same strategy has been successfully applied to enrich HSCs from mouse cyclophosphamide/granulocytecolony stimulating factor (G-CSF)-mobilized cells, 3 mouse fetal liver, 4 as well as from the BM of various strains of mice 5 and older mice. 3 Recent data examining the overlap between SLAM family member expression with the Hoechst dye efflux side population in conjunction with canonical HSC cell surface markers have confirmed the potential of CD150 ϩ selection for mouse HSC enrichment. 6 Although it has been suggested that some HSC activity may also be present in the CD150 Ϫ cell fraction of mouse cells, 6 data from multiple groups indicate that there is little or no long-term HSC activity in the CD150 Ϫ fraction of mouse hematopoietic cells. 1,3,4,[7][8][9][10] In humans, the CD34 cell surface marker is largely used in clinical applications for isolation of HSCs and progenitor cells, and as a predictor of graft HSC content in transplants. The combination CD34 ϩ CD38 Ϫ provides further enrichment (1 in 600 to 1 in 3500), 11 but the heterogeneity of this population precludes studies requiring levels of purity achieved in mouse models, such as comparing gene expression profiles or in situ histologic analyses. Given the use of SLAM receptors for identifying long-term repopulating HSCs in mice, 1,3,4,7-10 we hypothesized that this strategy may similarly facilitate the isolation of highly enriched populations of HSCs in humans and nonhuman p...

Wild-Type Measles Virus Interferes with Short-Term Engraftment of Human CD34 + Hematopoietic Progenitor Cells

“…Confirming a recent study (26), a minor subpopulation of CD34 ϩ HS/PCs expressing CD150 was detectable, which represent progenitor rather than stem cells. Wild-type MV infection of CD34 ϩ HS/PCs was inefficient, with little evidence of cell destruction.…”

“…SLAMF receptors are also involved in early phases of lineage commitment during hematopoiesis, and differential expression of CD150, CD48 (SLAMF2), and CD244 (SLAMF4) marks subsets of murine hematopoietic stem and progenitor cells (HS/PCs). This led to the term "SLAM code," whose applicability to human CD34 ϩ HS/PCs is as yet controversial (24,26,30,43,50).…”

Measles virus envelope pseudotyped lentiviral vectors transduce quiescent human HSCs at an efficiency without precedent

“…Of note, CD46 expression levels increased on cytokine-stimulated as compared with resting hCD34 1 cells, marked by a twofold increase in mean fluorescent intensity ( Figure 3A). In contrast, SLAM was almost undetectable at the surface of quiescent and stimulated hCD34 1 cells as reported, 31,33 which confirmed that SLAM, a valid surface marker for murine HSCs is not applicable for hHSCs ( Figure 3A and Larochelle et al 30 ). Nectin-4 was also not detected on HSCs.…”

“…28,29 This led to the term "SLAM code" whose applicability to human HSCs (hHSCs) and progenitors is still controversial. [28][29][30][31][32] Indeed, several reports already showed the absence of CD150 on the hCD34 …”

Markers for Hematopoietic Stem Cells: Histories and Recent Achievements

“…Although essential difference has not been observed between mouse and man regarding either the organs producing HSC or the transcription factors regulating their differentiation, completely different markers have been used to sort HSC in the two species. Human HSC do not express Sca1 or the SLAM family CD150 (Larochelle et al, 2011). While the CD34 + CD38 -phenotype has been regarded as the canonical marker set for human HSC, it has been repeatedly reported that murine adult HSC locate in the CD34 -CD38 + fraction (Randall et al, 1996;Matsuoka et al, 2001;Tajima et al, 2001).…”