![]() 8,9īreast cancer PDXs are initiated either orthotopically in the mammary fat pad(s) or heterotopically in subcutaneous space on the mid-dorsal flank(s) of immature female mice. There have been limited direct comparisons of receptiveness of strains, but cell line evidence suggests that NSG mice are more receptive to tumor development. Meric-Bernstam, in Patient Derived Tumor Xenograft Models, 2017 Implantation Techniquesīreast cancer PDXs have been generated in athymic nude, NOD-SCID, or NSG mice. Although this method is suitable for the detection of small, and perhaps insignificant, levels of chimerism, PCR data cannot be reliably used for quantification of the level of chimerism. When levels of engraftment are low, such that it cannot be detected by flow cytometric analysis, investigators have relied on polymerase chain reaction (PCR) analysis to detect human genes in the murine bone marrow. Alternatively, this approach has been used to deliver graft cells directly into the bone marrow microenvironment and avoid the need for homing of cells into the bone marrow of recipient animals. Ī technique suitable for multiple samplings of bone marrow cells from the femur can be used to temporally examine the kinetics of human engraftment.To reduce the impact of the reticuloendothelial system of NOD/SCID mice on transplanted human graft cells, injection of up to 10 × 10 7 nonadherent, irradiated CD34 − cells a few hours before transplantation or mixed with graft cells has been practiced. Use of antibiotics for a few days before and a few days after radiation was found to enhance survival and overall health of transplanted mice (E. These cytokines both protected recipient mice from the effects of radiation and modulated the behavior of chimeric human hematopoietic cells. Human cytokines can be used to supplement transplanted mice. In reverse, T cell engraftment in NOD/SCID mice is almost completely absent in most cases unless specific types of NOD/SCID mice are used. Human hematopoietic engraftment in NOD/SCID mice is preferentially skewed toward the B cell lineage, whereby multilineage assessment reveals a large percentage of B cells. Therefore monitoring engraftment in the blood of recipient mice over time may generate negative results. Įven well‐engrafted mice with high levels of chimerism in the bone marrow do not necessarily have detectable human cells in the periphery.Usually mice with >1% T cells are problematic. It is therefore critical to examine these mice for the presence of murine T cells in the periphery and to eliminate positive mice from the experimental design. ![]() ![]() These T cells can interfere with and possibly eliminate human engraftment. NOD/SCID mice are “leaky.” Therefore, some mice will develop mouse T cells that can be detected in the periphery as CD3 + cells. In this section, a few peculiarities of the NOD/SCID system are listed to enable investigators to avoid common problems with the use of this model in examining cord blood hematopoietic stem cells in vivo. ![]()
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