Recombinant IL-7/HGFβ efficiently induces transplantable murine hematopoietic stem cells

Difficulty obtaining sufficient hematopoietic stem cells (HSCs) directly from the donor has limited the clinical use of HSC transplantation. Numerous attempts to stimulate the ex vivo growth of purified HSCs with cytokines and growth factors generally have induced only modest increases in HSC numbers while decreasing their in vivo reconstituting ability. We previously developed a recombinant single-chain form of a naturally occurring murine hybrid cytokine of IL-7 and the beta chain of hepatocyte growth factor (rIL-7/HGF beta) that stimulates the in vitro proliferation and/or differentiation of common lymphoid progenitors, pre-pro-B cells, and hematopoietic progenitor cells (day 12 spleen colony-forming units) in cultures of mouse BM. Here we used the rIL-7/HGF beta in culture to induce large numbers of HSCs from multiple cell sources, including unseparated BM cells, purified HSCs, CD45(-) BM cells, and embryonic stem cells. In each instance, most of the HSCs were in the G(0) phase of the cell cycle and exhibited reduced oxidative stress, decreased apoptosis, and increased CXCR4 expression. Furthermore, when injected i.v., these HSCs migrated to BM, self-replicated, provided radioprotection, and established long-term hematopoietic reconstitution. These properties were amplified by injection of rIL-7/HGF beta directly into the BM cavity but not by treatment with rIL-7, rHGF, and/or rHGF beta.