AC133(+) G(0) cells from cord blood show a high incidence of long-term culture-initiating cells and a capacity for more than 100 million-fold amplification of colony-forming cells in vitro

AC133(+) cells may provide an alternative to CD34(+) cells as a target for cell expansion and gene therapy protocols. We examined the differences in proliferative potential between cord blood selected for AC133 or CD34 in serum-free, stroma cell-free culture for up to 30 weeks. Because most hemopoietic stem cells reside within the G(0)/G(1) phase of the cell cycle, we combined enrichment according to AC133 or CD34 expression with G(0) position in the cell cycle to identify populations enriched for putative stem cells. Our results show that AC133(+) G(0) cells demonstrated a long-term culture-initiating cell incidence of 1 in 4.2 cells, had a colony-forming cell incidence of 1 in 2.8 cells, were capable of producing 660 million-fold expansion of nucleated cells and 120 million-fold expansion of colony-forming units-granulocyte-macrophage over a period of 30 weeks, and were consistently superior to CD34(+) G(0) cells according to these parameters. Furthermore, we have shown that AC133(+)CD34(-) cells have the ability to generate CD34(+) cells in culture, which suggests that at least some AC133(+) cells are ancestral to CD34(+) cells. We conclude that AC133 isolation provides a better means of selection for primitive hemopoietic cells than CD34 and that, in combination with isolation according to G(0) phase of the cell cycle, AC133 isolation identifies a highly enriched population of putative stem cells.