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  

Construction and preliminary evaluation of a mouse subcutaneous chimeric model of renal vascular unit based on isolation of kidney-derived cells from human urine

作　　者：蒋超[1] 王超[1] 任晓楠[1] 周晓辉[1] JIANG Chao;WANG Chao;REN Xiaonan;ZHOU Xiaohui(Shanghai Public Health Clinical Center,Fudan University,Shanghai 201508,China)

机构地区：[1]复旦大学附属公共卫生临床中心,上海201508

出　　处：《中国实验动物学报》2022年第2期216-222,共7页Acta Laboratorium Animalis Scientia Sinica

基　　金：上海市科学技术委员会实验动物专项(19140905300,19140905301);上海市卫生健康委员会项目(20204Y0335);上海市公共卫生临床中心院级科研课题(KY-GW-2021-05,KY-GW-2021-38);上海实验动物安全控制管理信息平台研究及应用示范(21140900200);生物安全实验室实验动物信息可追溯系统应用评价研究(21140900202)。

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。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.

关 键 词：肾源细胞 人肾血管单位小鼠模型 皮下注射 间充质干细胞 人脐静脉内皮细胞 

分 类 号：Q95-33[生物学—动物学]