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中华细胞与干细胞杂志(电子版) ›› 2026, Vol. 16 ›› Issue (01) : 23 -30. doi: 10.3877/cma.j.issn.2095-1221.2026.01.003

论著

人类21三体(47,XX/XY,+21)诱导多能干细胞的制备
王承伟1, 李娟娟2,3, 胡文杰2,3, 陈致帆2,3, 许尹2,3,()   
  1. 1230051 合肥市妇幼保健院检验科
    2230032 合肥,安徽医科大学精神卫生与心理科学学院
    3230032 合肥,安徽医科大学分子神经精神病学实验室
  • 收稿日期:2025-05-27 出版日期:2026-02-01
  • 通信作者: 许尹
  • 基金资助:
    国家自然科学基金(82271465,81601107)

Generation of human trisomy 21 (47, XX/XY, +21) induced pluripotent stem cells

Chengwei Wang1, Juanjuan Li2,3, Wenjie Hu2,3, Zhifan Chen2,3, Yin Xu2,3,()   

  1. 1Department of Clinical Laboratory, Hefei Maternal and Child Health Hospital, 230051 Hefei, China
    2School of Mental Health and Psychological Sciences, Anhui Medical University, 230032 Hefei, China
    3Laboratory of Molecular Neuropsychiatry, Anhui Medical University, 230032 Hefei, China
  • Received:2025-05-27 Published:2026-02-01
  • Corresponding author: Yin Xu
引用本文:

王承伟, 李娟娟, 胡文杰, 陈致帆, 许尹. 人类21三体(47,XX/XY,+21)诱导多能干细胞的制备[J/OL]. 中华细胞与干细胞杂志(电子版), 2026, 16(01): 23-30.

Chengwei Wang, Juanjuan Li, Wenjie Hu, Zhifan Chen, Yin Xu. Generation of human trisomy 21 (47, XX/XY, +21) induced pluripotent stem cells[J/OL]. Chinese Journal of Cell and Stem Cell(Electronic Edition), 2026, 16(01): 23-30.

目的

建立唐氏综合征(DS)患者外周血单个核细胞(PBMCs)来源的诱导多能干细胞(iPSCs)模型,研究DS相关神经系统疾病。

方法

采用电转重编程技术将2例DS儿童的PBMCs重编程为iPSCs。通过形态学观察、核型分析、流式细胞术、免疫荧光染色和三胚层分化实验对获得的iPSCs进行系统表征,流式细胞术数据以阳性细胞百分比表示。

结果

成功获得具有典型形态的2株iPSCs (DS1-iPSCs和DS2-iPSCs),核型分析实验发现其核型为47,XX或XY,+21,证实2例患者均患有21-三体综合征。流式细胞术和免疫荧光检测结果显示iPSCs表达多能性标志物SSEA4、TRA-1-60、OCT4和SOX2。三胚层分化实验证实iPSCs可分化为内胚层(AFP、SOX17)、中胚层(α-SMA、BRACHYURY)和外胚层(Tuj1、PAX6)细胞,具有多向分化潜能。

结论

本研究成功建立2例DS患儿来源的iPSCs模型,细胞具有21-三体核型、多能性标志物表达及三胚层分化能力,为DS相关神经系统疾病研究提供新的细胞模型和研究工具。

Objective

To establish an induced pluripotent stem cells (iPSCs) model derived from peripheral blood mononuclear cells (PBMCs) of Down syndrom (DS) patients, providing a new research tool for studying DS and its associated neurological disorders.

Methods

The PBMCs of two children with DS were reprogrammed into iPSCs by using the electric reprogramming technology. The obtained iPSCs were systematically characterized by morphological observation, karyotype analysis, flow cytometry, immunofluorescence staining, and three-germ-layer differentiation experiments. Flow cytometry data are expressed as the percentage of positive cells.

Results

Two iPSCs lines, DS1-iPSCs and DS2-iPSCs, with typical morphology were successfully obtained. Karyotype analysis revealed a karyotype of 47,XX or XY,+21, confirming that both patients had Trisomy 21. Flow cytometry and immunofluorescence results showed that the iPSCs expressed pluripotency markers SSEA4, TRA-1-60, OCT4, and SOX2. Three-germ-layer differentiation experiments confirmed that the iPSCs could differentiate into endoderm (AFP, SOX17) , mesoderm (α-SMA, BRACHYURY) , and ectoderm (Tuj1, PAX6) cells, demonstrating their pluripotency.

Conclusion

This study successfully established iPSCs model derived from the peripheral blood of two DS children, which possess trisomy 21 karyotype, expression of pluripotency markers and the ability to differentiate from the three germ layers, offering a new cell model and research tool for investigating DS and its associated neurological disorders.

图1 在全自动荧光显微镜EVOS明场下观察DS1-iPSCs和DS2-iPSCs细胞形态注:a图为患者1诱导的iPSCs;b图为患者2诱导的iPSCs
图2 DS1-iPSCs和DS2-iPSCs核型鉴定注:a图为DS1-iPSCs核型鉴定为47条染色体(XX,+21);b图为DS2-iPSCs核型鉴定为47条染色体(XY,+21)
图3 DS1-iPSCs和DS2-iPSCs流式鉴定多能性注:a图为DS1-iPSCs流式鉴定多能干细胞标志物SSEA4;b图为DS1-iPSCs流式鉴定多能干细胞标志物Tra-1-60;c图为DS2-iPSCs流式鉴定多能干细胞标志物SSEA4;d图为DS2-iPSCs流式鉴定多能干细胞标志物Tra-1-60
图4 全自动荧光显微镜EVOS荧光RFP,GFP和DAPI通道下观察DS1-iPSCs和DS2-iPSCs多能干细胞标志物的表达注:鉴定DS1-iPSC和DS2-iPSC表达多能干细胞标志物OCT4、SOX2
图5 全自动荧光显微镜EVOS荧光RFP,GFP和DAPI通道下观察DS1-iPSCs和DS2-iPSCs三胚层分化标志物的表达注:鉴定DS1-iPSC和DS2-iPSC向胚层分化后表达内胚层标志物AFP、SOX17、中胚层标志物α-SMA、Brachyury、外胚层标志物Tuj1、PAX6
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