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AC133⁺ G₀ 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

Cancer Research UK Department of Experimental Haematology, Paterson Institute for Cancer Research, Manchester, United Kingdom

Key Words. AC133⁺ cells • CD34⁺ cells • Cord blood • Ex vivo expansion • G₀/G₁ phase of cell cycle

Correspondence: Dr. Yvonne J. Summers, Department of Medical Oncology, Christie Hospital NHS Trust, Manchester, M20 4BX United Kingdom. Telephone: 0161-446-3741; Fax: 0161-446-3299; e-mail: yvonnejsummers{at}aol.com

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 G₀/G₁ phase of the cell cycle, we combined enrichment according to AC133 or CD34 expression with G₀ position in the cell cycle to identify populations enriched for putative stem cells. Our results show that AC133⁺ G₀ 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⁺ G₀ 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 G₀ phase of the cell cycle, AC133 isolation identifies a highly enriched population of putative stem cells.

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