Human CD34-negative hematopoietic stem cells: The current understanding of their biological nature

•Human cord blood-derived CD34-negative HSCs are identified and characterized.•CD133 GPI-80 specific positive/enrichment markers for CD34– HSCs.•CD34–HSCs constitute class of human different from CD34+ reside at the apex HSC hierarchy.•Proposal A revised road map is proposed commitment CB-derived CD34–HSCs. Hematopoietic stem cell (HSC) heterogeneity hierarchy a current topic interest, having major implications clinical transplantation basic research on HSCs. It was long believed that most primitive in mammals, including mice humans, were CD34 antigen positive (CD34+). However, 2 decades ago, it reported murine long-term multilineage reconstituting lineage marker negative (Lin–, i.e., c-kit+Sca-1+CD34low/–), known as CD34low/– KSL cells. In contrast, HSCs, counterpart cells, hard to identify time mainly because their rarity. We previously very blood (CB)-derived severe combined immunodeficiency (SCID)-repopulating cells (SRCs) using intra-bone marrow injection method new concept SRCs (HSCs) hierarchy. Through series studies, we two markers: CD133 GPI-80. The combination these enabled development an ultrahigh-resolution purification well successful both single-cell level. Cell population purity crucial prerequisite reliable biological molecular analyses. Clonal analyses highly purified have revealed potent megakaryocyte/erythrocyte differentiation potential. Based observations, propose This review updates understanding nature Fluorescence-activated sorting (FACS) technology has markedly progressed over past 3 decades. 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