Sustained Multilineage Engraftment of Allogeneic Hematopoietic Stem Cells in NOD/SCID Mice After In Utero Transplantation

Abstract

Substantial barriers exist to the engraftment of hematopoietic cells in mice after in utero transplantation. Although high levels of donor-derived hematopoiesis have been reported in SCID mice, the majority of chimeric recipients exhibit decreasing levels of donor cells over time. To directly test whether the natural killer cell and macrophage activity of the recipients represents a barrier to sustained engraftment, fetal NOD/SCID mice were injected on day 13.5 of gestation with an enriched congenic hematopoietic progenitor cell population. Forty-four percent of pups showed the presence of Ly5.1+ donor cells 4 weeks after transplantation. The mean number of donor-derived nucleated cells in the peripheral blood (PB) was 30%. Although the majority of circulating donor cells were lymphocytes, up to 15% expressed myelomonocytic markers. Serial PB samples from individual mice indicated that the percentage of circulating donor cells increased from 17% to 55% between 4 and 24 weeks. At 6 months posttransplantation, an increased frequency of multilineage, donor-derived cells was also observed in the bone marrow (BM) and the spleen of chimeric recipients. The engraftment of pluripotent hematopoietic stem cells was evaluated by transplanting BM from chimeric mice into irradiated congenic recipients. Irradiated secondary recipients also exhibited multilineage donor-derived hematopoiesis in the PB, BM, and spleen for up to 6 months. These results show that the in utero transplantation of lineage-depleted BM cells into NOD/SCID recipients produces a high frequency of sustained engraftment of allogeneic hematopoietic stem cells.

SCID Mouse Models of Human Stem Cell Engraftment

“…A second approach may be the in utero injection of human stem cells into NOD-scid mice. It has recently been observed that in utero injection of allogeneic cells into day 13-14 gestation NOD-scid fetuses results in engraftment of the allogeneic stem cells in the absence of irradiation [120]. If the NOD-Rag1 null mice support enhanced human cell engraftment similar to that observed in NOD-scid mice, or if human stem cells can be engrafted in utero without the need for irradiation, two of the major limitations of the hu-SRC-NOD-SCID model would be overcome.…”

In utero transplantation of human fetal haemopoietic cells in NOD/SCID mice

“…Of a total of 65 embryos in seven pregnant dams 51 (78%) were injected with human FL cells and 33 (51%) were live‐born. We directly compared these results to those obtained when we transplanted allogeneic lineage‐negative haemopoietic progenitor cells into NOD/SCID recipients in a parallel series of experiments ( Archer et al , 1997 ). Of a total of 84 embryos, in nine pregnant dams, 53 (63%) were injected and 36 pups (43%) were born.…”

“…High levels of donor cell engraftment in both mice and humans have been limited to situations in which the recipient has a stem cell or lineage‐specific defect ( Archer et al , 1997 ; Blazar et al , 1995a , b; Howson‐Jan et al , 1993 ; Fleischman & Mintz, 1979). If the defect inhibits stem cell function as in W v mice, enhanced engraftment of normal donor stem cells would be the expected result.…”

“…The NOD/SCID strain demonstrates a high degree of immune dysfunction which correlates with high frequencies of engraftment and high levels of donor cells following injection of human cells into adult mice ( Cashman et al , 1997 ; Larochelle et al , 1996 ; Bhatia et al , 1997 ; Wang et al , 1997 ; Greiner et al , 1995 ; Lowry et al , 1996 ). We have recently shown that when enriched murine haemopoietic progenitor cells are injected into NOD/SCID mice in utero , significant levels of sustained multilineage engraftment are found in up to 44% of the recipients ( Archer et al , 1997 ). These studies indicate that the NOD/SCID mouse supports the growth and differentiation of both xenogeneic and allogeneic haemopoietic progenitor cells.…”

In utero transplanted human hepatocytes allow postnatal engraftment of human hepatocytes in pigs

“…[14][15][16][17] Previously, the in utero transplantation of hematopoietic cells led to stable long-term engraftment in the postnatal environment. 14,[16][17][18] In this study, we show for the first time that IUCT in piglets with adult human hepatocytes allows the stable postnatal engraftment of human hepatocytes. The effectiveness of our IUCT step was determined by a cytotoxicity assay that measured the change in piglet serum reactivity to human PBLs after 3 postnatal SQ priming injections using human hepatocytes.…”