从人体尿液中分离肾源细胞构建拟肾血管单位小鼠皮下嵌合模型及其初步评价
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复旦大学附属公共卫生临床中心,上海 201508

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Construction and preliminary evaluation of a mouse subcutaneous chimeric model of renal vascular unit based on isolation of kidney-derived cells from human urine
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Shanghai Public Health Clinical Center, Fudan University, Shanghai 201508, China

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    摘要:

    目的 将健康人体尿液中分离培养的肾源细胞与人源间充质干细胞( mesenchymal stem cells, MSCs)、人脐静脉内皮细胞( human umbilical vein endothelial cells,HUVECs)以 3 ∶3 ∶2的比例混合,并移植到 NOD- SCID 免疫缺陷小鼠皮下而构建人源化嵌合小鼠模型。 方法 (1)体外培养 MSCs、HUVECs,同时从健康人体尿液中分离培养肾源细胞,并对该细胞进行免疫荧光鉴定。 (2)将上述 3 种细胞以 3 ∶ 3 ∶ 2 的比例混合后与 Matrigel 基质胶一起注射到 NOD-SCID 小鼠皮下形成人源组织,通过 HE 染色、免疫组织化学对移植模型进行鉴定。 结果 (1)成功地从尿液中分离培养出肾源细胞,并检测鉴定了肾小管上皮细胞和集合管细胞;(2)将 3 种细胞混合后注射到 NOD-SCID 免疫缺陷小鼠皮下可初步形成小管结构以及与小鼠血管相连通的人源血管结构。 结论 具有肾小管上皮细胞和集合管细胞的肾源细胞与 HUVECs 和 MSCs 皮下移植入 NOD-SCID 免疫缺陷小鼠体内可形成嵌合的人肾血管单位小鼠模型。该模型可为肾病个体化治疗和相关病毒感染研究提供一个人源化动物模型。

    Abstract:

    Objective To construct a humanized chimeric mouse model by subcutaneously transplanting NOD- SCID immunodeficient mice with kidney-derived cells cultured from healthy human urine mixed with human mesenchymal stem cells (MSCs) and human umbilical vein endothelial cells ( HUVECs). Methods ( 1) Kidney-derived cells were isolated from healthy human urine and cultured in vitro. MSCs and HUVECs were also cultured in vitro. All three kinds of cells were identified by immunofluorescence. (2)The three cell types were then mixed in a ratio of 3 ∶ 3 ∶ 2 and injected subcutaneously with Matrigel into NOD-SCID mice to develop human tissue. The transplantation model was identified by hematoxylin and eosin staining and immunohistochemistry. Results (1) Kidney-derived cells were successfully isolated and cultured from urine, and renal tubular epithelial cells and collecting duct cells were detectable and identified by immunohistochemistry. (2) After subcutaneous injection into NOD-SCID immunodeficient mice, the three kinds of cells initially developed tubule structures and human-derived vascular structures that communicated with the murine systemic circulation. Conclusions We successfully created a chimeric mouse model of the human renal vascular unit by subcutaneous transplantation of kidney-derived cells, HUVECs, and MSCs, including renal tubular epithelial cells and collecting tubular cells, into NOD-SCID immunodeficient mice. This model may provide a humanized animal model for research into the individualized treatment of human chronic kidney disease and kidney-related viral infections.

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蒋超,王超,任晓楠,周晓辉.从人体尿液中分离肾源细胞构建拟肾血管单位小鼠皮下嵌合模型及其初步评价[J].中国实验动物学报,2022,30(2):216~222.

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  • 收稿日期:2022-01-06
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  • 在线发布日期: 2022-06-27
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