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首页> 外文期刊>Stem Cell Studies >Detection of human hematopoietic stem cell engraftment in the livers of adult immunodeficient mice by an optimized flow cytometric method
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Detection of human hematopoietic stem cell engraftment in the livers of adult immunodeficient mice by an optimized flow cytometric method

机译：优化流式细胞仪检测成年免疫缺陷小鼠肝脏中人造血干细胞的植入

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Immunodeficient NOD.Cg-Prkdcscid Il2rgtm1Wjl/ SzJ (NSG) mice are a valuable resource to study human hematopoietic stem cells. Prolonged multilineage hematopoiesis indicates stem cell engraftment and generally is measured by flow cytometry. In this study, we took advantage of the multi-parameter detection afforded by modern flow cytometers to optimize detection of human hematopoiesis in NSG mice. Antigens widely expressed by mouse or human cells were evaluated as markers to distinguish mixtures of these cells to optimize and test the limits of chimerism detection. The bone marrow, spleen, and liver of NSG mice transplanted with human hematopoietic cells were analyzed for evidence of engraftment. Mouse bone marrow cells were best marked for exclusion by staining with a combination of CD45, TER-119, and anti-H-2Kd monoclonal antibodies, whereas live human cells were most accurately identified by elimination of cell doublets and positive staining for CD59. Human stem cells (CD34++CD133+CD38low) and progenitors were detected in the bone marrow and liver, but not in the spleen. An unusual pattern of myeloid antigen expression was detected in the bone marrow and CD3+CD4+CD8+ T-cells were detected in the spleen. We concluded that multicolor flow cytometric analysis that clearly distinguishes mouse and human cells offers accurate detection of human chimerism in NSG mice. Human hematopoiesis can be detected in the bone marrow and liver of NSG mice with T-lymphopoiesis, possibly occurring in the spleen.
机译：免疫缺陷的NOD.Cg-Prkdcscid Il2rgtm1Wjl / SzJ（NSG）小鼠是研究人类造血干细胞的宝贵资源。延长的多谱系造血表明干细胞植入，通常通过流式细胞术进行测量。在这项研究中，我们利用了现代流式细胞仪提供的多参数检测来优化NSG小鼠中人类造血功能的检测。评价了小鼠或人类细胞广泛表达的抗原作为标记，以区分这些细胞的混合物，从而优化和测试嵌合体检测的极限。分析了移植有人类造血细胞的NSG小鼠的骨髓，脾脏和肝脏，以寻找植入的证据。通过用CD45，TER-119和抗H-2Kd单克隆抗体的组合染色可以最好地标记小鼠骨髓细胞是否被排斥，而通过消除细胞双峰和CD59阳性染色可以最准确地鉴定活人细胞。在骨髓和肝脏中检测到人干细胞（CD34 ++ CD133 + CD38low）和祖细胞，但在脾脏中未检测到。在骨髓中检测到异常的髓样抗原表达模式，在脾脏中检测到CD3 + CD4 + CD8 + T细胞。我们得出的结论是，可以清楚地区分小鼠和人类细胞的多色流式细胞仪分析可准确检测NSG小鼠中的人类嵌合体。可以在患有T淋巴细胞生成的NSG小鼠的骨髓和肝脏中检测到人类造血功能，这可能发生在脾脏中。

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