中国组织工程研究

中国组织工程研究 ›› 2026, Vol. 30 ›› Issue (7): 1720-1729.doi: 10.12307/2026.089

• 脐带脐血干细胞 umbilical cord blood stem cells • 上一篇    下一篇

高活性脐带间充质干细胞对衰老树鼩胸腺结构和功能的影响

叶倩倩1，2，潘  杭2，田  川2，朱向情2，叶  丽2，赵晓娟2，舒莉萍1，潘兴华2   

  1. 1贵州医科大学、基础医学院免疫学教研室、组织工程与干细胞实验中心，贵州省贵阳市   550004；2中国人民解放军联勤保障部队第九二〇医院干细胞与免疫细胞生物医药技术国家地方联合工程实验室、云南省细胞治疗技术转化医学重点实验室、昆明市干细胞与再生医学重点实验室、基础医学实验室，云南省昆明市   650032
  • 收稿日期:2025-01-09 修回日期:2025-06-13 接受日期:2025-07-03 出版日期:2026-03-08 发布日期:2025-08-19
  • 通讯作者: 潘兴华，博士，主任医师，中国人民解放军联勤保障部队第九二〇医院干细胞与免疫细胞生物医药技术国家地方联合工程实验室、云南省细胞治疗技术转化医学重点实验室、昆明市干细胞与再生医学重点实验室、基础医学实验室，云南省昆明市 650032 共同通讯作者：舒莉萍，博士，教授，贵州医科大学、基础医学院免疫学教研室、组织工程与干细胞实验中心，贵州省贵阳市 550004
  • 作者简介:叶倩倩，女，2000年生，贵州省遵义市人，汉族，贵州医科大学在读硕士，主要从事脐带间充质干细胞抗衰老研究。
  • 基金资助:
    云南省细胞治疗技术医学重点实验室(2015DG034)，项目负责人：潘兴华；云南省科技计划项目重大科技专项(2018ZF007)，项目负责人：潘兴华

Effects of highly active umbilical cord mesenchymal stem cells on structure and function of thymus in elderly tree shrews

Ye Qianqian1, 2, Pan Hang2, Tian Chuan2, Zhu Xiangqing2, Ye Li2, Zhao Xiaojuan2, Shu Liping1, Pan Xinghua2   

  1. 1Center for Tissue Engineering and Stem Cell Experiment, Department of Immunology, College of Basic Medical Sciences, Guizhou Medical University, Guiyang 550004, Guizhou Province, China; 2National and Local Joint Engineering Laboratory of Stem Cell and Immune Cell Biomedical Technology, 920 Hospital of China People’s Liberation Army Joint Logistics Support Force, Yunnan Provincial Key Laboratory of Cell Therapy Technology Translational Medicine, Kunming Key Laboratory of Stem Cell and Regenerative Medicine, Basic Medical Laboratory, Kunming 650032, Yunnan Province, China
  • Received:2025-01-09 Revised:2025-06-13 Accepted:2025-07-03 Online:2026-03-08 Published:2025-08-19
  • Contact: Pan Xinghua, PhD, Chief physician, National and Local Joint Engineering Laboratory of Stem Cell and Immune Cell Biomedical Technology, 920 Hospital of China People’s Liberation Army Joint Logistics Support Force, Yunnan Provincial Key Laboratory of Cell Therapy Technology Translational Medicine, Kunming Key Laboratory of Stem Cell and Regenerative Medicine, Basic Medical Laboratory, Kunming 650032, Yunnan Province, China; Shu Liping, MD, Professor, Center for Tissue Engineering and Stem Cell Experiment, Department of Immunology, College of Basic Medical Sciences, Guizhou Medical University, Guiyang 550004, Guizhou Province, China
  • About author:Ye Qianqian, Master candidate, Center for Tissue Engineering and Stem Cell Experiment, Department of Immunology, College of Basic Medical Sciences, Guizhou Medical University, Guiyang 550004, Guizhou Province, China; National and Local Joint Engineering Laboratory of Stem Cell and Immune Cell Biomedical Technology, 920 Hospital of China People’s Liberation Army Joint Logistics Support Force, Yunnan Provincial Key Laboratory of Cell Therapy Technology Translational Medicine, Kunming Key Laboratory of Stem Cell and Regenerative Medicine, Basic Medical Laboratory, Kunming 650032, Yunnan Province, China
  • Supported by:
     Medical Key Laboratory of Cell Therapy Technology of Yunnan Province, No. 2015DG034 (to PXH); Major Science and Technology Special Project of Yunnan Provincial Science and Technology Plan Project, No. 2018ZF007 (to PXH)

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摘要：

文题释义：

高活性脐带间充质干细胞：课题组前期在脐带间充质干细胞中发现了一群比现有脐带间充质干细胞体积小、生长活性更高的亚群，暂命名为高活性脐带间充质干细胞。已发现高活性脐带间充质干细胞对猕猴卵巢、胸腺衰老及树鼩脾脏衰老与全身多器官功能障碍综合征等多种动物模型的治疗效果显著。
胸腺衰老：胸腺是人体最先发生衰老的器官，表现为胸腺萎缩及免疫功能下降，因此延缓胸腺衰老有利于提高免疫功能。

摘要
背景：胸腺作为免疫系统核心器官，是T淋巴细胞分化、发育、成熟的场所。在青春期时，胸腺就开始发生萎缩退化，是人类最早出现衰老的器官。目前干预胸腺衰老的研究仍较少，高活性脐带间充质干细胞改善树鼩胸腺衰老研究国内尚未见报道。
目的：探讨高活性脐带间充质干细胞对衰老树鼩胸腺结构和功能的影响。
方法：通过剖腹产获取新生树鼩脐带组织，使用组织贴壁法分离培养获得高活性脐带间充质干细胞，使用吉凯基因绿色荧光蛋白以感染复数值140共培养72 h为转染条件，转染第4代高活性脐带间充质干细胞。20只平均年龄在7周岁的雌性树鼩，随机分为老年模型组与老年治疗组，每组10只；10只年龄在3周岁的雌性树鼩设为青年对照组。老年治疗组通过尾静脉输注第4代高活性脐带间充质干细胞，剂量为1×107个/kg，1次/d，连续3次；青年对照组与老年模型组不做任何特殊处理。常规饲养4个月取胸腺组织，苏木精-伊红染色观察胸腺组织结构；Masson染色观察胸腺组织纤维化程度；免疫组织化学染色检测衰老标志分子p21、p53表达；免疫荧光染色检测增殖蛋白Ki67、增殖细胞核抗原表达；活性氧荧光染色检测胸腺组织的活性氧含量；免疫组织化学染色检测胸腺组织CD3+总T淋巴细胞数量；酶联免疫吸附实验检测树鼩血清中胸腺素β4水平；DAPI复染胸腺组织细胞核观察高活性脐带间充质干细胞在胸腺组织的分布情况。

结果与结论：①老年治疗组树鼩胸腺中观察到吉凯基因绿色荧光蛋白转染的高活性脐带间充质干细胞；②与老年模型组相比，老年治疗组树鼩胸腺紊乱结构得到明显改善，胸腺实质组织增加(P < 0.05)，脂肪减少，皮髓质界限清晰，胶原纤维占比降低，具有向年轻状态变化趋势；③与老年模型组相比，老年治疗组衰老标志分子p21、p53表达下调(P < 0.01，P < 0.05)；④与老年模型组相比，老年治疗组增殖与凋亡分子Ki67表达呈增加趋势，但差异无显著性意义(P > 0.05)，增殖细胞核抗原表达增加(P < 0.05)，活性氧水平表达显著下降(P < 0.01)；⑤与老年模型组相比，老年治疗组CD3+T淋巴细胞比例呈增加趋势，但差异无显著性意义(P > 0.05)；⑥与老年模型组相比，老年治疗组血清胸腺素β4水平增加(P < 0.01)；⑦研究结果表明，高活性脐带间充质干细胞能够改善衰老树鼩胸腺组织结构，降低衰老标志分子表达水平，提高胸腺细胞的增殖活性和胸腺功能。

https://orcid.org/0009-0007-8433-695X (叶倩倩) 


中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程


关键词: 干细胞, 脐带间充质干细胞, 树鼩, 胸腺, 衰老, 衰老标志分子, 增殖基因, 免疫

Abstract: BACKGROUND: As the central organ of the immune system, the thymus serves as the primary site for T lymphocyte differentiation, development and maturation. Thymic involution initiates during adolescence, making it the first organ to undergo age-related degeneration in humans. There is still few current research on therapeutic interventions for thymic aging, and there are no studies in China addressing the potential of highly active umbilical cord mesenchymal stem cells to modulate thymic aging in the tree shrew. 
OBJECTIVE: To investigate the effects of highly active umbilical cord mesenchymal stem cells on the structure and function of thymus of aging tree shrews.
METHODS: The umbilical cord tissues from newborn tree shrews were obtained via cesarean section, and highly active umbilical cord mesenchymal stem cells were isolated and cultured using the tissue adherence method. Fourth-passage highly active umbilical cord mesenchymal stem cells were transfected using green fluorescent protein from GeneChip under conditions of a multiplicity of infection of 140 for 72 hours. Twenty female tree shrews with an average age of 7 years were randomly divided into elderly model group and elderly treatment group (10 per group). Ten 3-year-old female tree shrews were designated as the young control group. The aged treatment group received tail vein infusions of fourth-passage highly active umbilical cord mesenchymal stem cells at a dose of 1×10⁷ cells/kg, once daily for three consecutive days. The young control group and aged model group received no special treatment. After 4 months of routine feeding, thymus tissues were collected. Hematoxylin-eosin staining was used to examine thymic structure. Masson staining was utilized to assess thymic fibrosis. Immunohistochemical staining was applied to detect the expression of senescence markers p21 and p53. Immunofluorescence staining was employed to evaluate the expression of proliferative proteins Ki67 and proliferating cell nuclear antigen. Reactive oxygen species fluorescence staining was used to measure reactive oxygen species levels in thymic tissues. Immunohistochemical staining was utilized to quantify CD3+ total T lymphocyte counts in the thymus. Enzyme-linked immunosorbent assay was employed to determine serum thymosin β4 levels in tree shrews. DAPI counterstaining of thymic nuclei was performed to observe the distribution of highly active umbilical cord mesenchymal stem cells in thymic tissues. 
RESULTS AND CONCLUSION: (1) Green fluorescent protein from GeneChip transfected tree shrew highly active umbilical cord mesenchymal stem cells were observed in the tree shrew thymus of the elderly treatment group. (2) Compared with the elderly model group, the thymus of the elderly treatment group exhibited significantly improved histoarchitecture, including increased thymic parenchyma (P < 0.05), reduced adipose infiltration, distinct corticomedullary demarcation, decreased collagen fiber proportion, and a trend toward rejuvenation. (3) Senescence markers p21 and p53 expression levels were downregulated in the elderly treatment group compared with the elderly model group (P < 0.01 and P < 0.05). (4) Compared with the elderly model group, the elderly treatment group showed a non-significant increasing trend in Ki67 expression (P > 0.05), elevated proliferating cell nuclear antigen expression (P < 0.05), and significantly reduced reactive oxygen species levels (P < 0.01). (5) Compared with the elderly model group, the proportion of CD3+ T lymphocytes displayed an increasing trend in the elderly treatment group, though the difference was not statistically significant (P > 0.05). (6) Serum thymosin β4 levels were significantly increased in the elderly treatment group compared with the elderly model group (P < 0.01). (7) These findings demonstrate that highly active umbilical cord mesenchymal stem cells ameliorate age-related thymic structural degeneration, suppress senescence marker expression, enhance thymocyte proliferative activity and thymic function in aged tree shrews.

Key words: stem cells, umbilical cord mesenchymal stem cells, tree shrew, thymus, aging, aging marker molecule, proliferating gene, immunity

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