邢义高,张磊磊,吕玲,桑宵,韩浩,高峰,雒猛

• 1:山东省细胞组织库
• 2:山东省齐鲁干细胞工程有限公司
• 3:山东省脐带血造血干细胞库

摘要(Abstract)：

目的 探讨保存时间对人脐带间充质干细胞来源外泌体物理稳定性的影响。方法培养人脐带间充质干细胞并鉴定表面标志物，观察第3代人脐带间充质干细胞的成骨、成脂和成软骨诱导分化能力。分离并制备人脐带间充质干细胞来源外泌体，观察外泌体的形态，检测外泌体的颗粒数、粒径、蛋白含量以及表面标志物的蛋白表达，并评估保存时间对外泌体物理稳定性的影响。结果人脐带间充质干细胞高表达CD90、CD44、CD73和CD105，低表达CD45和人类白细胞抗原-DR。人脐带间充质干细胞在成骨、成脂、成软骨诱导分化后，分别观察到红色的钙结节、红色的圆形脂滴、蓝色和立体的圆球状结构。外泌体的粒径峰值为（121.56±1.27）nm，颗粒数为（2.13±0.12）×10~(11)/mL，蛋白含量为（0.71±0.03）mg/mL。外泌体形态为杯状或茶托形，且具有双层膜结构。外泌体表达肿瘤易感基因101(TSG101)、CD9、CD81和CD63蛋白，不表达钙连蛋白。随着保存时间的延长，外泌体的形态无明显变化，外泌体的粒径和蛋白含量亦无明显变化（P>0.05）。与保存0个月比较，保存5、6个月时外泌体颗粒数、TSG101和CD81蛋白表达减少（P<0.05）。结论 保存时间对外泌体形态、粒径、蛋白含量无明显影响，而对颗粒数、CD81和TSG101的表达有明显影响。在进行外泌体研究时，应选择保存时间在5个月以内的外泌体样本。

关键词(KeyWords)： 人脐带间充质干细胞;外泌体;保存时间

Abstract:

Keywords:

基金项目(Foundation):

作者(Author): 邢义高,张磊磊,吕玲,桑宵,韩浩,高峰,雒猛

DOI: 10.19460/j.cnki.0253-3685.2025.09.001

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