Enhanced Reconstitution of Human Erythropoiesis and Thrombopoiesis in an Immunodeficient Mouse Model with Kit Wv Mutations

Abstract

SummaryIn human-to-mouse xenograft models, reconstitution of human hematopoiesis is usually B-lymphoid dominant. Here we show that the introduction of homozygous KitWv mutations into C57BL/6.Rag2nullIl2rgnull mice with NOD-Sirpa (BRGS) strongly promoted human multi-lineage reconstitution. After xenotransplantation of human CD34+CD38− cord blood cells, these newly generated C57BL/6.Rag2nullIl2rgnullNOD-Sirpa KitWv/Wv (BRGSKWv/Wv) mice showed significantly higher levels of human cell chimerism and long-term multi-lineage reconstitution compared with BRGS mice. Strikingly, this mouse displayed a robust reconstitution of human erythropoiesis and thrombopoiesis with terminal maturation in the bone marrow. Furthermore, depletion of host macrophages by clodronate administration resulted in the presence of human erythrocytes and platelets in the circulation. Thus, attenuation of mouse KIT signaling greatly enhances the multi-lineage differentiation of human hematopoietic stem and progenitor cells (HSPCs) in mouse bone marrow, presumably by outcompeting mouse HSPCs to occupy suitable microenvironments. The BRGSKWv/Wv mouse model is a useful tool to study human multi-lineage hematopoiesis.

A Highly Efficient and Faithful MDS Patient-Derived Xenotransplantation Model for Pre-Clinical Studies

“…Very little is known about the causes of the phenotypic heterogeneity in MDS and genotype-phenotype studies would greatly advance our mechanistic understanding of this complex entity. To date, immunodeficient mouse models have supported erythro-and megakaryopoiesis solely from fetal liverand cord blood-derived HSPCs 40,41 , further enhanced by mutation of the murine ckit receptor conferring impaired function to murine stem cells [42][43][44] and likely murine erythropoietic progenitors 45 . None of these models have supported erythro-and megakaryopoiesis from adult HSPCs.…”

Epithelial‐mesenchymal transition is activated in CD44‐positive malignant ascites tumor cells of gastrointestinal cancer

“…Quantitative PCR (qPCR) was performed using the StepOnePlus Real‐time PCR System. Multiplex qPCR of ATC and primary tissues was performed using the TaqMan PreAmp Master Mix (Applied Biosystems, Foster City, CA, USA), and cDNA samples were subjected to qPCR using the BioMark system (Fluidigm, San Francisco, CA, USA) as previously reported . Reactions were run in triplicate in 3 independent experiments.…”

“…(Applied Biosystems, Foster City, CA, USA), and cDNA samples were subjected to qPCR using the BioMark system (Fluidigm, San Francisco, CA, USA) as previously reported. 16 Reactions were run in triplicate in 3 independent experiments. The geometric mean of the housekeeping gene GAPDH was used as the internal control.…”

Enhanced differentiation of functional human T cells in NSGW41 mice with tissue-specific expression of human interleukin-7

“…NSGW41 mice carry the hypomorph W41 allele in the Kit gene, harbor the NOD-specific variant of the Sirpa gene, are T-, B- and NK-cell deficient based on null mutations in Prkdc and Il2rg genes, and allow for human donor stem cell engraftment in the absence of preconditioning 2, 3 . NSGW41 mice that are stably engrafted with human hematopoietic stem cell display continuous human hematopoiesis with increased myeloid 2, 4 , megakaryocytic and erythroid output compared to irradiated NSG recipient mice 5, 6 .…”