中国组织工程研究

中国组织工程研究 ›› 2026, Vol. 30 ›› Issue (19): 4853-4859.doi: 10.12307/2026.674

• 脂肪干细胞 adipose-derived stem cells • 上一篇    下一篇

人脂肪多系分化应激耐受细胞治疗大鼠缺血性脑卒中

高洪梅1，张  坤1，2，肖东杰1，2，刘  华1，2   

  1. 1山东第一医科大学附属中心医院细胞治疗中心，山东省济南市  250013；2山东省移植与组织工程技术研究中心，山东省济南市  250013
  • 收稿日期:2025-05-26 接受日期:2025-09-18 出版日期:2026-07-08 发布日期:2026-02-13
  • 通讯作者: 刘华，博士，主任技师，山东第一医科大学附属中心医院细胞治疗中心，山东省济南市 250013；山东省移植与组织工程技术研究中心，山东省济南市 250013
  • 作者简介:高洪梅，女，1973年生，2004年青岛大学毕业，硕士，副主任技师，主要从事分子诊断相关工作。
  • 基金资助:
    济南市科技发展计划(201907053)，项目负责人：刘华

Human adipose multilineage-differentiating stress-enduring cells on treatment of ischemic stroke in rats

Gao Hongmei1, Zhang Kun1, 2, Xiao Dongjie1, 2, Liu Hua1, 2   

  1. 1Cell Therapy Center, Central Hospital Affiliated to Shandong First Medical University, Jinan 250013, Shandong Province, China; 2Shandong Research Center of Transplantation and Tissue Engineering Technology, Jinan 250013, Shandong Province, China
  • Received:2025-05-26 Accepted:2025-09-18 Online:2026-07-08 Published:2026-02-13
  • Contact: Liu Hua, MD, Senior technologist, Cell Therapy Center, Central Hospital Affiliated to Shandong First Medical University, Jinan 250013, Shandong Province, China; Shandong Research Center of Transplantation and Tissue Engineering Technology, Jinan 250013, Shandong Province, China
  • About author:Gao Hongmei, MS, Associate senior technologist, Cell Therapy Center, Central Hospital Affiliated to Shandong First Medical University, Jinan 250013, Shandong Province, China
  • Supported by:
    Jinan Science and Technology Development Plan, No. 201907053 (to LH)

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摘要：

文题释义：

缺血性脑卒中：是由于脑的供血动脉突然阻塞，导致大脑特定区域血流中断，引起脑部组织不可逆损伤，是全球范围内常见的脑血管疾病，具有高发病率、高复发率、高致残率、高死亡率及高经济负担的“五高”特点。
多系分化应激耐受细胞：即Muse细胞，为SSEA3+CD105+成体干细胞，在多个组织器官中广泛存在，正常机体含量极少。该细胞具有三胚层分化能力，可迁移到受损部位，耐受恶劣环境，还具有自发分化为归巢组织相容细胞的能力。

摘要
背景：间充质干细胞治疗缺血性脑卒中取得了良好的效果，从脂肪间充质干细胞中分离获得的多系分化应激耐受(multilineage-differentiating stress-enduring，Muse)细胞在缺血性脑卒中的研究尚少。
目的：探讨脂肪Muse细胞静脉移植对大鼠缺血性脑卒中的神经修复作用。
方法：胰酶长时间处理(4 h)人脂肪间充质干细胞，通过磁珠分选获得阶段特异性胚胎抗原3阳性细胞(Muse细胞)。制备大鼠大脑中动脉阻塞模型，造模成功后，脂肪间充质干细胞组、脂肪Muse组经尾静脉注射200 μL脂肪间充质干细胞悬液或脂肪Muse细胞悬液(含2×105细胞)，生理盐水组注射200 μL生理盐水。移植后第3，7天进行大鼠行为学评分，移植后第3天苏木精-伊红染色观察脑组织损伤情况，免疫荧光检测脑组织损伤区微管相关蛋白2及Ki67表达，TUNEL染色观察脑组织损伤区细胞凋亡情况，Western blot检测脑组织损伤区生长相关蛋白43及Bcl-2、Bax蛋白表达。
结果与结论：①磁珠分选后，流式细胞仪检测显示阳性分选细胞的阶段特异性胚胎抗原3表达率高达80%；②细胞移植后第7天，与生理盐水组相比，脂肪间充质干细胞组与脂肪Muse细胞组神经功能损伤评分降低(P < 0.05)；细胞移植后第3天，与脂肪间充质干细胞组相比，脂肪Muse细胞组神经功能损伤评分降低(P < 0.05)；③苏木精-伊红染色观察到脂肪间充质干细胞组与脂肪Muse细胞组大鼠脑皮质区炎症反应及空泡形成减轻，微管相关蛋白2荧光染色显示脂肪Muse细胞组抑制神经元丢失更为明显；④TUNEL染色及Ki67荧光染色结果显示脂肪间充质干细胞组与脂肪Muse细胞组损伤区细胞凋亡减少，细胞增殖增多；⑤Western blot结果显示脂肪间充质干细胞组与脂肪Muse细胞组Bcl-2/Bax比值、生长相关蛋白43蛋白表达升高，脂肪Muse细胞组在抑制细胞凋亡方面优于脂肪间充质干细胞组。结果表明，脂肪间充质干细胞与脂肪Muse细胞移植大鼠神经修复良好，脂肪Muse细胞在抑制细胞凋亡方面发挥更好的作用。

关键词: 脂肪间充质干细胞, Muse细胞, 缺血性脑卒中, 阶段特异性胚胎抗原3(SSEA3), 多系分化应激耐受细胞

Abstract: BACKGROUND: Mesenchymal stem cells have shown good therapeutic effects in ischemic stroke, while the role of multilineage-differentiating stress-enduring (Muse) cells isolated from adipose-derived mesenchymal stem cells in ischemic stroke needs further study. 
OBJECTIVE: To explore the neurorestorative effect of intravenous administration of adipose Muse cells on ischemic stroke in rats.
METHODS: The Muse cells expressing stage-specific embryonic antigen 3 were sorted by magnetic beads after long-term (4 hours) trypsin incubation of human adipose-derived mesenchymal stem cells. The middle cerebral artery occlusion model was established in rats. After successful modeling, the adipose-derived mesenchymal stem cell group and adipose-derived Muse group were injected with 200 μL adipose-derived mesenchymal stem cell suspension or adipose-derived Muse cell suspension (containing 2×105 cells) via the tail vein. The saline group was injected with 200 μL saline. Behavioral scores of rats were evaluated at 3 and 7 days after injection. At 3 days after injection, hematoxylin-eosin staining was used to detect brain tissue damage. Immunofluorescence was used to detect the expression of microtubule-associated protein 2 and Ki67 in the brain tissue injury area. TUNEL staining was used to observe cell apoptosis in the injury area. Western blot assay was used to detect the protein expression of growth-associated protein 43, Bcl2, and Bax. 
RESULTS AND CONCLUSION: (1) After magnetic bead sorting, the flow cytometry showed that the expression rate of stage-specific embryonic antigen 3 was as high as 80%. (2) At 7 days after injection, the neurological function injury scores of the adipose mesenchymal stem cell group and the adipose Muse cell group were reduced compared with those of the saline group (P < 0.05). At 3 days after cell transplantation, the neurological function injury scores of the adipose Muse cell group were reduced compared with those of the adipose mesenchymal stem cell group (P < 0.05). (3) Hematoxylin-eosin staining showed that the inflammatory response and vacuolation in the cerebral cortex of rats in the adipose mesenchymal stem cell group and the adipose Muse cell group were alleviated. Microtubule-associated protein 2 fluorescence staining showed that the adipose Muse cell group inhibited neuronal loss more significantly. (4) TUNEL staining and Ki67 fluorescence staining results showed that apoptosis of cells in the injured area of the adipose mesenchymal stem cell group and the adipose Muse cell group was reduced, and cell proliferation was increased. (5) Western blot assay results showed that the Bcl-2/Bax ratio and growth-associated protein 43 protein expression in the adipose mesenchymal stem cell group and the adipose Muse cell group were increased, and the adipose Muse cell group was superior to the adipose mesenchymal stem cell group in inhibiting cell apoptosis. The results showed that the nerves of rats transplanted with adipose mesenchymal stem cells and adipose Muse cells were well repaired, and Muse cells played a better role in inhibiting cell apoptosis.

Key words: adipose mesenchymal stem cell, Muse cell, ischemic stroke, stage-specific embryonic antigen 3 (SSEA3), multilineage-differentiating stress-enduring cells

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