A revised road map for the commitment of human cord blood CD34-negative hematopoietic stem cells

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Abstract
We previously identified CD34-negative (CD34−) severe combined immunodeficiency (SCID)-repopulating cells as primitive hematopoietic stem cells (HSCs) in human cord blood. In this study, we develop a prospective ultra-high-resolution purification method by applying two positive markers, CD133 and GPI-80. Using this method, we succeed in purifying single long-term repopulating CD34−HSCs with self-renewing capability residing at the apex of the human HSC hierarchy from cord blood, as evidenced by a single-cell-initiated serial transplantation analysis. The gene expression profiles of individual CD34+and CD34−HSCs and a global gene expression analysis demonstrate the unique molecular signature of CD34−HSCs. We find that the purified CD34−HSCs show a potent megakaryocyte/erythrocyte differentiation potential in vitro and in vivo. Megakaryocyte/erythrocyte progenitors may thus be generated directly via a bypass route from the CD34−HSCs. Based on these data, we propose a revised road map for the commitment of human CD34−HSCs in cord blood.

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