EGF-responsive human fetal NSC-derived BM cells are able to establish long-term hematopoietic reconstitution in SCID-hu mice.

One million EGF-responsive human fetal NSCs were injected directly into each human bone graft in SCID-hu mice, and human BM cells were harvested from those bone fragments 4 months after injection. One million of those harvested human BM cells were then directly injected into each human graft, including bone and thy/liv, in SCID-hu mice, and graft cells were harvested 4 months later and subjected to flow cytometry for donor-derived hematopoietic cells. (A) Intrathymic T-cell development of EGF-generated primary spheres. Graft cells were analyzed for T-cell markers CD3, CD4, and CD8, and donor marker HLA-MA2.1. The percentage of T cells expressing detectable levels of donor-specific class I antigen was recorded. (B) B-cell and myeloid-cell differentiation of EGF-generated primary spheres in implanted human fetal bone fragments. Graft cells were analyzed for B-cell marker CD19 and myeloid marker CD33, and a donor marker for EGF-generated primary spheres (HLA-MA2.1–positive). The percentage of B and myeloid cells expressing detectable levels of donor-specific class I antigen was recorded.