Bone marrow reconstitution of NOD/SCID mice Patients provided consent preoperatively for use of tissue obtained at surgery. The study was approved by the Research Ethics Board of the University Health Network (UHN, Toronto, Ontario, Canada). Red blood cells (RBC) were removed and CD34+ HSPC enriched by negative selection using the StemSep system (StemCell Technologies, Vancouver, BC, Canada). Cells and bone fragments were diluted in phosphate buffered saline containing sodium citrate and 2% fetal calf serum, shaken for 30 min and filtered through a cell strainer. MATERIALS AND METHODS Patient samples and CD34+ cell enrichmentīone marrow was obtained from patients with OA who were undergoing total hip replacement. As proof of principle we used HSPC from patients with osteoarthritis (OA) to address the feasibility of using HSPC from adult patients to repopulate the immune system of NOD/SCID recipient mice. Our hypothesis was that immune events leading to development of human arthritis can be recapitulated in NOD/SCID mice by transfer of HSPC.
NOD SCID MOUSE FULL
Nonobese diabetic/severe combined immunodeficiency (NOD/SCID) mice have been shown to accept human HSPC and may allow the engraftment of the full spectrum of immunocompetent cells. Hematopoietic stem and progenitor cells (HSPC) are pluripotent cells that are capable of repopulating the immune system and thereby reestablishing the immune repertoire of the donor. Yet animal models that faithfully recapitulate the cellular events of these diseases have also proved difficult to develop. Bone marrow-derived human HSPC engraft NOD/SCID mice and traffic appropriately to an inflammatory stimulus in the joint, thus offering the potential for direct studies on the immunopathogenesis and treatment of human arthritis.Ī rigorous analysis of the immune basis of inflammatory joint diseases in patients has been limited by ethical and technical constraints. Chlamydia-injected mice that had been repopulated with HSPC had synovial inflammation, consisting of human neutrophils and macrophages.Ĭonclusion. Human B lymphoid and myeloid cells were detected in the bone marrow and spleen 6 weeks following transfer of HSPC, and engrafted recipient mice remained healthy up to 12 weeks postinjection. Human bone marrow HSPC successfully engrafted NOD/SCID mice, with some mice showing up to 90% engrafted human cells. Arthritis was induced by an intraarticular injection of Chlamydia trachomatis and injected knee joints were examined 5 days later by histology and immunohistochemistry. Human cell engraftment was analyzed by flow cytometry. HSPC were enriched by negative selection and injected into the femur of irradiated anti-CD122 treated nonobese diabetic/severe combined immunodeficiency (NOD/SCID) mice. Bone marrow was obtained from patients with osteoarthritis who were undergoing total hip replacement. Our hypothesis is that it is possible to develop humanized arthritis models through novel techniques of hematopoietic stem and progenitor cell (HSPC) delivery. Studies of human inflammatory arthritis would be significantly aided by the development of better animal models.
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