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Constitutively Active -Catenin Promotes Expansion of Multipotent Hematopoietic Progenitors in Culture¹

Yoshihiro Baba^*, Takafumi Yokota, Hergen Spits, Karla P. Garrett^*, Shin-Ichi Hayashi and Paul W. Kincade^2,*

^* Immunobiology and Cancer Program, Oklahoma Medical Research Foundation, Oklahoma City, OK 73104; Department of Hematology and Oncology, Osaka University, Suita, Osaka, Japan; Department of Cell Biology and Histology of the Academic Medical Center, University of Amsterdam, Amsterdam, The Netherlands; and Department of Molecular and Cellular Biology, School of Life Science, Faculty of Medicine, Tottori University, Yonago, Tottori, Japan

This study was designed to investigate one component of the Wnt/-catenin signaling pathway that has been implicated in stem cell self-renewal. Retroviral-mediated introduction of stable -catenin to primitive murine bone marrow cells allowed the expansion of multipotential c-Kit^lowSca-1^low/–CD19^– CD11b/Mac-1^–Flk-2^–CD43⁺AA4.1⁺NK1.1^–CD3^–CD11c^–Gr-1^–CD45R/B220⁺ cells in the presence of stromal cells and cytokines. They generated myeloid, T, and B lineage lymphoid cells in culture, but had no T lymphopoietic potential when transplanted. Stem cell factor and IL-6 were found to be minimal requirements for long-term, stromal-free propagation, and a -catenin-transduced cell line was maintained for 5 mo with these defined conditions. Although multipotential and responsive to many normal stimuli in culture, it was unable to engraft several types of irradiated recipients. These findings support previous studies that have implicated the canonical Wnt pathway signaling in regulation of multipotent progenitors. In addition, we demonstrate how it may be experimentally manipulated to generate valuable cell lines.

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