Page 17 - 南京医科大学自然科学版第1期

第41卷第1期 南京医科大学学报（自然科学版）
2021年1月 Journal of Nanjing Medical University（Natural Sciences） · 11 ·

·基础研究·

人多能干细胞诱导类肾的实验研究

俞 欣 ，李开霖 ，孔 峰 ，赵升田 ，张登禄 4*
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中国科学技术大学附属第一医院（安徽省立医院）急诊外科，安徽 合肥 230001；山东大学第二医院中心实验室，山东 济
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南 250000；山东省立医院山东省泌尿器官与功能重建工程实验室，山东 济南 250000；山东中医药大学附属医院实验中
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心，山东 济南 250000
［摘 要］ 目的：体外诱导人多能干细胞（human pluripotent stem cell，hPSC）来源的类肾体，探讨类肾的结构成熟度、血管化和
非肾系分化现象。方法：通过肾上皮分化试剂盒诱导hPSC成类肾，用免疫荧光观察类肾元结构中特异性标志物CDH1、LTL和
PODX的表达，透射电镜（transmission electron microscopy，TEM）观察肾小球和肾小管微观结构；第15天类肾移植到SCID小鼠
肾包膜下，4周后处死动物，取移植物行组织形态学观察，免疫荧光法观察移植物中人抗核抗原（human nuclear antigen，HNA）、
CDH1、PODX和MECA⁃32的表达。结果：肾脏特异性标志物CDH1、LTL和PODX呈阳性表达，电镜观察到不成熟的足细胞和
肾小管结构；移植后的类肾体积增大，观察到肉眼可见的血管，HE染色移植物内可见红细胞和类软骨组织，HNA、CDH1、PODX
和MECA⁃32呈阳性表达。结论：hPSC可体外诱导结构不成熟的类肾，在体内环境下，类肾可完成血管化并持续生长，但存在大量
非肾系结构。
［关键词］ 人多能干细胞；类肾；血管化
［中图分类号］ R329 ［文献标志码］ A ［文章编号］ 1007⁃4368（2021）01⁃011⁃06
doi：10.7655/NYDXBNS20210103

An experimental study on kidney organoids induced by human pluripotent stem cells

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YU Xin ，LI Kailin ，KONG Feng ，ZHAO Shengtian ，ZHANG Denglu 4*
1 Department of Emergency Surgery，the First Affiliated Hospital of USTC，Anhui Provincial Hospital，Division of
Life Science and Medicine，University of Science and Technology of China，Hefei 230001；Central Research Lab，the
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Second Hospital of Shandong University，Jinan 250000；Engineering Laboratory of Urinary Organ and Functional
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Reconstruction of Shandong Province，Shandong Provincial Hospital，Jinan 250000；Central Laboratory，Affiliated
Hospital of Shandong University of Traditional Chinese Medicine，Jinan 250000，China
［Abstract］ Objective：To investigate the maturity，vascularization and nonrenal differentiation of kidney organoids using induced
renal organoids derived human pluripotent stem cells（hPSCs）. Methods：hPSCs were induced into renal organoids cells through a
renal epithelial cell differentiation kit. The renal organoids structures were highly expressed with specific kidney markers CDH1，LTL，
and PODXl by immunofluorescence method. The microstructures of glomeruli and tubules were observed by transmission electron
microscopy；The animals were sacrificed under the renal capsule of SCID mice for 4 weeks，the grafts were taken for histomorphological
observation，and the expressions of HNA，CDH1，PODX and MECA⁃32 were observed by immunofluorescence method. Results：The
kidney markers CDH1，LTL，and PODX were positively expressed. Immature podocytes and tubular structures were observed by
transmission electron microscopy. The kidney organoid volume increased after transplantation，blood vessels were visible to the naked
eye，and HE stained grafts were visible red blood cells and cartilage⁃like tissues，HNA，CDH1，PODX and MECA⁃32 were positively
expressed. Conclusion：hPSC can induce immature kidney organoids structures in vitro. In vivo，renal orgaoids structures can complete
vascularization and continue to grow，but there were a large number of non⁃renal structures.
［Key words］ human pluripotent stem cell；renal organoid；vascularization
［J Nanjing Med Univ，2021，41（01）：011⁃015，053］

［基金项目］ 国家自然科学基金（81700592）
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通信作者（Corresponding author），E⁃mail：915489139@qq.com