Xenotransplantation is frequently used to study normal and malignant hematopoiesis of human cells. However, conventional mouse xenotransplantation models lack essential human-specific bone-marrow (BM)-microenvironment-derived survival, proliferation, and self-renewal signals for engraftment of normal and malignant blood cells. As a consequence, many human leukemias and other hematologic disorders do not robustly engraft in these conventional models. Here, we describe a complete workflow for the generation of humanized ossicles with an accessible BM microenvironment that faithfully recapitulates normal BM niche morphology and function. The ossicles, therefore, allow for accelerated and superior engraftment of primary patient-derived acute myeloid leukemia (AML) and other hematologic malignancies such as myelofibrosis (MF) in mice. The humanized ossicles are formed by in situ differentiation of BM-derived mesenchymal stromal cells (MSCs). Human hematopoietic cells can subsequently be transplanted directly into the ossicle marrow space or by intravenous injection. Using this method, a humanized engraftable BM microenvironment can be formed within 6-10 weeks. Engraftment of human hematopoietic cells can be evaluated by flow cytometry 8-16 weeks after transplantation. This protocol describes a robust and reproducible in vivo methodology for the study of normal and malignant human hematopoiesis in a more physiologic setting.

中文翻译：

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用于小鼠造血异种移植的人源化骨髓小骨龛的产生和使用。

异种移植经常用于研究人类细胞的正常和恶性造血。然而，传统的小鼠异种移植模型缺乏必要的人类特异性骨髓 (BM)-微环境衍生的生存、增殖和自我更新信号，用于植入正常和恶性血细胞。因此，许多人类白血病和其他血液系统疾病并没有在这些传统模型中牢固地植入。在这里，我们描述了一个完整的工作流程，用于生成具有可访问的 BM 微环境的人性化听小骨，该微环境忠实地概括了正常的 BM 生态位形态和功能。因此，听小骨可以加速和更好地植入原发性患者衍生的急性髓性白血病 (AML) 和其他血液系统恶性肿瘤，例如小鼠骨髓纤维化 (MF)。人源化听小骨由 BM 来源的间充质基质细胞 (MSCs) 原位分化形成。随后可以将人类造血细胞直接移植到听小骨骨髓空间或通过静脉注射。使用这种方法，可以在 6-10 周内形成人性化的可移植 BM 微环境。移植后 8-16 周可通过流式细胞术评估人类造血细胞的植入情况。该协议描述了一种强大且可重复的体内方法，用于在更生理的环境中研究正常和恶性人类造血。可在6-10周内形成人性化的可移植BM微环境。移植后 8-16 周可通过流式细胞术评估人类造血细胞的植入情况。该协议描述了一种强大且可重复的体内方法，用于在更生理的环境中研究正常和恶性人类造血。可在6-10周内形成人性化的可移植BM微环境。移植后 8-16 周可通过流式细胞术评估人类造血细胞的植入情况。该协议描述了一种强大且可重复的体内方法，用于在更生理的环境中研究正常和恶性人类造血。