中国组织工程研究

中国组织工程研究 ›› 2018, Vol. 22 ›› Issue (17): 2686-2691.doi: 10.3969/j.issn.2095-4344.0525

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

第2代和第5代人脐带间充质干细胞外泌体miRNA谱的比较

林庆铿1，2，尹德宏2，刘菊芬2，张学娟1，2，白盈盈1，2，宋乙甲1，2，潘兴华2，刘高米洋2   

  1. 1昆明医科大学解放军昆明总医院临床学院，云南省昆明市 650032；2解放军昆明总医院细胞生物治疗中心，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，云南省昆明市 650032
  • 修回日期:2018-04-17 出版日期:2018-06-18 发布日期:2018-06-18
  • 通讯作者: 刘高米洋，硕士，解放军昆明总医院细胞生物治疗中心，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，云南省昆明市 650032
  • 作者简介:林庆铿，男，1986年生，福建省福州市连江县人，汉族，昆明医科大学在读硕士，主要从事人脐带间充质干细胞与衰老相关研究。 并列第一作者：尹德宏，主治医师，主要从事脊髓损伤及干细胞治疗方面的研究。
  • 基金资助:

    解放军动物专项(SYDW(2016)004)；云南省重点实验室(2015DG034)

Exosomes from human umbilical cord mesenchymal stem cells at passage 2 and 5: a comparative study on microRNA profiles

Lin Qing-keng1, 2, Yin De-hong2, Liu Ju-fen2, Zhang Xue-juan1, 2, Bai Ying-ying1, 2, Song Yi-jia1, 2, Pan Xing-hua2, Liu-Gao Mi-yang2   

  1. 1Kunming General Hospital Clinical College of Kunming Medical University, Kunming 650032, Yunnan Province, China; 2Cell Biological Therapy Center of Kunming General Hospital of PLA, Cell Biological Medicine integrated Engineering Laboratory of State and Region of Yunnan Province, the Stem Cell Therapy Key Laboratory of Yunnan Province, Kunming 650032, Yunnan Province, China
  • Revised:2018-04-17 Online:2018-06-18 Published:2018-06-18
  • Contact: Liu-Gao Mi-yang, Master, Cell Biological Therapy Center of Kunming General Hospital of PLA, Cell Biological Medicine integrated Engineering Laboratory of State and Region of Yunnan Province, the Stem Cell Therapy Key Laboratory of Yunnan Province, Kunming 650032, Yunnan Province, China
  • About author:Lin Qing-keng, Master candidate, Kunming General Hospital Clinical College of Kunming Medical University, Kunming 650032, Yunnan Province, China; Cell Biological Therapy Center of Kunming General Hospital of PLA, Cell Biological Medicine integrated Engineering Laboratory of State and Region of Yunnan Province, the Stem Cell Therapy Key Laboratory of Yunnan Province, Kunming 650032, Yunnan Province, China. Yin De-hong, Attending physician, Cell Biological Therapy Center of Kunming General Hospital of PLA, Cell Biological Medicine integrated Engineering Laboratory of State and Region of Yunnan Province, the Stem Cell Therapy Key Laboratory of Yunnan Province, Kunming 650032, Yunnan Province, China. Lin Qing-keng and Yin De-hong contributed equally to this work.
  • Supported by:

    the Animal Special Fund of PLA, No. SYDW(2016)004; the Key Laboratory Project of Yunnan Province, No. 2015DG034

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被引次数

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摘要：

文章快速阅读：

文题释义：
外泌体：是一类由某些细胞释放的细胞外囊泡。已有报道认为它们能够调节宿主-病原体的相互作用，参与传染性和炎性疾病、神经疾病和癌症等很多种疾病的病理过程，同时在正常的生理过程中也发挥着介导细胞间通讯等作用。另外细胞外囊泡含有丰富的生物标志物，可用于监测临床状态、治疗反应、疾病进展等，同时它们还有发展成临床药物递送载体的潜力。由于细胞外囊泡的组成成分并不是随机的，每一个细胞外囊泡都会携带特定的分子信息，如携带蛋白质、脂质、核酸和糖等生物活性分子在细胞间传递信号，其有一个复杂的分选系统来决定哪些分子能够进入细胞外囊泡，因而其有望成为进一步了解人体细胞层面调节过程的突破口。 
聚类分析：是通过建立各种不同的数学模型，把基于相似数据特征的变量或样本组合在一起。归为一个簇的 miRNA在功能上可能相似或关联，从而找到miRNA的功能信息。聚类分析将表达模式相同或相近的miRNA或样本聚集成类，并使用不同颜色的区域代表不同的聚类分组信息，从而判断不同样品或不同实验条件下的聚类模式。由于同类的miRNA可能具有相似的功能，或是共同参与同一代谢过程或细胞通路，因此有助于筛选潜在的有协同作用或受同样调控的miRNA，从而帮助发掘miRNA的功能。

 

摘要
背景：研究发现外泌体具有部分间充质干细胞的功能，了解间充质干细胞外泌体中发挥代表性作用的物质，可以为进一步探索人工合成外泌体类似物提供线索。
目的：研究第2代和第5代人脐带间充质干细胞来源外泌体中miRNA的表达谱差异。
方法：采用超高速离心法提取第2代和第5代人脐带间充质干细胞培养上清液中的外泌体，并使用高通量测序技术，对建立的文库进行测序，再将结果进行分析，了解不同外泌体之间所表达的miRNA，并做聚类分析。
结果与结论：①第2代人脐带间充质干细胞来源外泌体能够检测出427 657种miRNA，占检出总miRNA的68.93%；第5代人脐带间充质干细胞来源外泌体能够检测出119 283种miRNA，占检出总miRNA的19.22%，其中二者共有的miRNA有73 526种，占检出总 miRNA的11.85%；②经生物信息学分析(聚类分析)表明这些miRNA可能参与了161种生物过程，包括细胞修复、免疫、抗衰老等；③随着传代次数的增加，外泌体miRNA成分发生了较大的变化，部分miRNA拷贝数水平明显减低，其中含量最高的前5个miRNA(has-miR-146a-5p、has-miR-191-5p、has-miR-493-3p、has-miR-423-5p、has-miR-134-5p)有望做为合成外泌体类似物的组成部分。

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
ORCID: 0000-0002-1883-560X(林庆铿)

关键词: 脐带间充质干细胞, 外泌体, microRNA, 高通量测序, 聚类分析, 干细胞

Abstract:

BACKGROUND: Exosomes have the function of some mesenchymal stem cells. Understanding the substance composition that plays a representative role in mesenchymal stem cell exosomes will provide clues for further exploration of synthetic exosome analogues.
OBJECTIVE: To investigate the difference of microRNA expression profiles in exosomes derived from passage 2 and 5 human umbilical cord mesenchymal stem cells (hUC-MSCs).
METHODS: Exosomes in the supernatant of passage 2 and 5 hUC-MSCs were extracted by ultra-high speed centrifugation. The established library was sequenced by using high-throughput sequencing technology. Then we analyzed the sequence results so as to understand the microRNA expression between different groups, and finally did a cluster analysis.
RESULTS AND CONCLUSION: 427 657 kinds of microRNAs were detected in the exosomes from passage 2 hUC-MSCs, accounting for 68.93% of the total microRNAs detected; and 119283 microRNAs were detected in the exosomes from passage 5 hUC-MSCs, accounting for 19.22% of the total microRNAs detected. There were 73 526 microRNAs shared between the exosomes from passage 2 and passage 5 hUC-MSCs, accounting for 11.85% of the total microRNAs detected. Bioinformatics analysis (cluster analysis) results showed that these miRNAs were likely to be involved in 161 biological processes, including cell repair, immune and anti-aging. The microRNAs in exosomes from passage 2 to passage 5 hUC-MSCs were largely different. Partial miRNAs exhibited significantly reduced copy numbers. The top five microRNAs with a higher amount, including has-miR-146a-5p, has-miR-191-5p, has-miR-493-3p, has-miR-423-5p, and has-miR-134-5p, have the potential to be the component of synthetic exosome analogues.

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

Key words: Umbilical Cord, Mesenchymal Stem Cells, Exosomes, MicroRNAs, High-Throughput Screening Assays, Tissue Engineering

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