Generation of Mouse-Zebrafish Hematopoietic Tissue Chimeric Embryos for Hematopoiesis and Host-Pathogen Interaction Studies
Generation of Mouse-Zebrafish Hematopoietic Tissue Chimeric Embryos for Hematopoiesis and Host-Pathogen Interaction Studies
ABSTRACT Xenografts of the hematopoietic system are extremely useful as disease models and for translational research. Zebrafish xenografts have been widely used to monitor blood cancer cell dissemination and homing due to the optical clarity of embryos and larvae, which allow unrestricted in vivo visualization of migratory events. To broaden the scope of xenotransplantation studies in zebrafish, we have developed a technique that transiently generates hematopoietic tissue chimeras by transplanting murine bone marrow cells into zebrafish blastulae. This procedure leads to mammalian cell integration into the fish developmental hematopoietic program. Monitoring zebrafish chimeras at different time points post fertilization using in vivo time-lapse and confocal imaging showed murine cell co-localization with developing primitive and definitive hematopoietic tissues, intravasation into fish circulation, and dynamic hematopoietic cell-vascular endothelial and hematopoietic cell-niche interactions. Immunohistochemistry assays performed in chimeric animals showed that, after engraftment, murine cells expressed antigens related to i) hematopoietic stem and progenitor cells, ii) active cell proliferation, and iii) myeloid cell lineages. Lastly, xenografted zebrafish larvae infected with Klebsiella pneumoniae showed murine immune cells trafficking to bacterial foci and interacting with bacterial cells. Overall, these results show that mammalian bone marrow cells xenografted in zebrafish integrate into the host hematopoietic system revealing highly conserved molecular mechanisms of hematopoiesis between zebrafish and mammals. In addition, this procedure introduces a useful and simple method that improves and broadens the scope of hematopoietic tissue xenotransplantation studies in zebrafish.
Allende Miguel L.、Clatworthy Anne、Ernst Christoph、Parada-Kusz Margarita、Villablanca Eduardo J.、Jijon Humberto、Riquelme Raquel、Penaranda Cristina、Zon Leonard I.、Henninger Jonathan E.、Li Brian、Hung Deborah、Nair Anil V.、Hagedorn Elliott J.
Center for Genome Regulation, Facultad de Ciencias, Universidad de Chile||Department of Cell Biology and Human Anatomy. University of CaliforniaBroad Institute of MIT and Harvard||Department of Molecular Biology, Massachusetts General Hospital||Center for Computational and Integrative Biology, Massachusetts General Hospital||Department of Genetics, Harvard Medical SchoolBroad Institute of MIT and Harvard||Department of Molecular Biology, Massachusetts General Hospital||Center for Computational and Integrative Biology, Massachusetts General Hospital||Department of Genetics, Harvard Medical SchoolCenter for Genome Regulation, Facultad de Ciencias, Universidad de ChileImmunology and Allergy, Department of Medicine, Solna, Karolinska Institute and University HospitalGastrointestinal Research Group, Faculty of Medicine, University of CalgaryStem Cell Program and Division of Hematology/Oncology, Boston Children?ˉs Hospital and Dana-Farber Cancer Institute, Howard Hughes Medical Institute, Harvard Stem Cell Institute, Harvard Medical SchoolBroad Institute of MIT and Harvard||Department of Molecular Biology, Massachusetts General Hospital||Center for Computational and Integrative Biology, Massachusetts General Hospital||Department of Genetics, Harvard Medical SchoolStem Cell Program and Division of Hematology/Oncology, Boston Children?ˉs Hospital and Dana-Farber Cancer Institute, Howard Hughes Medical Institute, Harvard Stem Cell Institute, Harvard Medical SchoolStem Cell Program and Division of Hematology/Oncology, Boston Children?ˉs Hospital and Dana-Farber Cancer Institute, Howard Hughes Medical Institute, Harvard Stem Cell Institute, Harvard Medical SchoolStem Cell Program and Division of Hematology/Oncology, Boston Children?ˉs Hospital and Dana-Farber Cancer Institute, Howard Hughes Medical Institute, Harvard Stem Cell Institute, Harvard Medical SchoolBroad Institute of MIT and Harvard||Department of Molecular Biology, Massachusetts General Hospital||Center for Computational and Integrative Biology, Massachusetts General Hospital||Department of Genetics, Harvard Medical SchoolCenter for Systems Biology, Program in Membrane Biology, Division of Nephrology, Massachusetts General Hospital and Harvard Medical SchoolStem Cell Program and Division of Hematology/Oncology, Boston Children?ˉs Hospital and Dana-Farber Cancer Institute, Howard Hughes Medical Institute, Harvard Stem Cell Institute, Harvard Medical School
基础医学生物科学研究方法、生物科学研究技术分子生物学
ZebrafishXenotransplantationLive imagingHematopoiesisCell migrationHostpathogen interactions
Allende Miguel L.,Clatworthy Anne,Ernst Christoph,Parada-Kusz Margarita,Villablanca Eduardo J.,Jijon Humberto,Riquelme Raquel,Penaranda Cristina,Zon Leonard I.,Henninger Jonathan E.,Li Brian,Hung Deborah,Nair Anil V.,Hagedorn Elliott J..Generation of Mouse-Zebrafish Hematopoietic Tissue Chimeric Embryos for Hematopoiesis and Host-Pathogen Interaction Studies[EB/OL].(2025-03-28)[2025-08-02].https://www.biorxiv.org/content/10.1101/216895.点此复制
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