Adult Murine Bone Marrow-Derived Very Small Embryonic-Like Stem Cells (VSELs) Differentiate Into the Hematopoietic Lineage After Co-Culture Over OP9 Stromal Cells

Abstract

Objective: We recently identified a population of small Sca-1(+)/Lin(-)/CD45(-) cells in adult murine bone marrow that express several epiblast/germ line and pluripotent stem cell markers (e.g., Oct-4 and SSEA-4) that we named "very small embryonic-like stem cells" (VSELs). In this report, we test the hypothesis that VSELs can differentiate along the hemato/lymphopoietic lineage. Materials and methods: Purified from bone marrow, VSELs were primed/cocultured over OP9 stroma cell line and subsequently tested in vitro and in vivo assays for their hematopoietic potential. In parallel, cells derived from VSELs were evaluated for expression of hematopoietic genes and surface markers. Results: Although we observed that freshly isolated VSELs do not exhibit in vitro and in vivo hematopoietic potential, they may, after coculture over OP9 stromal cells, differentiate along the hematopoietic lineage in a similar way as embryonic stem cells or inducible pluripotent stem cells. "OP9-primed," VSEL-derived cells acquired expression of several hemato/lymphopoiesis-specific genes and markers, gave rise to hematopoietic colonies in vitro, and protected lethally irradiated mice in both primary and secondary transplant models on transplantation. We also observed that, compared to hematopoietic stem/progenitor cells, VSELs are highly resistant to total body irradiation. Conclusions: Based on these observations, we postulate that VSELs are the most primitive murine bone marrow-residing population of stem cells that have the potential to become specified into the hematopoietic lineage and may share some of the characteristics of long-term repopulating HSCs.