Isolation and Comprehensive Analysis of Cochlear Tissue-Derived Small Extracellular Vesicles

被引：3

作者：

Jiang, Pei ^{[1
,2
]}

Ma, Xiangyu ^{[1
]}

Wang, Xinlin ^{[1
]}

Huang, Jingyuan ^{[1
]}

Wang, Yintao ^{[1
]}

Ai, Jingru ^{[1
,2
]}

Xiao, Hairong ^{[1
,2
]}

Dai, Mingchen ^{[1
]}

Lin, Yanqin ^{[1
,2
]}

Shao, Buwei ^{[3
]}

Tang, Xujun ^{[1
]}

Tong, Wei ^{[1
]}

Ye, Zixuan ^{[1
,2
]}

Chai, Renjie ^{[1
,4
,5
,6
]}

Zhang, Shasha ^{[1
]}

机构：

[1] Southeast Univ, Zhongda Hosp, Jiangsu Prov High Tech Key Lab Biomed Res, State Key Lab Digital Med Engn,Dept Otolaryngol He, Nanjing 210096, Peoples R China

[2] Southeast Univ, Shenzhen Res Inst, Shenzhen 1518063, Peoples R China

[3] Tel Aviv Univ, Fac Med & Hlth Sci, Sch Med, IL-6997801 Tel Aviv, Israel

[4] Univ Elect Sci & Technol China, Sichuan Prov Peoples Hosp, Dept Otolaryngol Head & Neck Surg, Chengdu 610072, Peoples R China

[5] Nantong Univ, Coinnovat Ctr Neuroregenerat, Nantong 226001, Peoples R China

[6] Chinese Acad Sci, Inst Stem Cell & Regenerat, Beijing 100101, Peoples R China

来源：

ADVANCED SCIENCE | 2024年 / 11卷 / 48期

基金：

中国国家自然科学基金;

关键词：

cochlea; FGFR1; hair cells; miRNA; small extracellular vesicle; supporting cells; CELL-PROLIFERATION; FGFR1; ACTIVATION; SURVIVAL; GROWTH; BIOMARKERS; MIGRATION; PATHWAY;

D O I：

10.1002/advs.202408964

中图分类号：

O6 [化学];

学科分类号：

0703 ;

摘要：

Small extracellular vesicles (sEVs) act as a critical mediator in intercellular communication. Compared to sEVs derived from in vitro sources, tissue-derived sEVs can reflect the in vivo signals released from specific tissues more accurately. Currently, studies on the role of sEVs in the cochlea have relied on studying sEVs from in vitro sources. This study evaluates three cochlear tissue digestion and cochlear tissue-derived sEV (CDsEV) isolation methods, and first proposes that the optimal approach for isolating CDsEVs using collagenase D and DNase & Iukcy; combined with sucrose density gradient centrifugation. Furthermore, it comprehensively investigates CDsEV contents and cell origins. Small RNA sequencing and proteomics are performed to analyze the miRNAs and proteins of CDsEVs. The miRNAs and proteins of CDsEVs are crucial for maintaining normal auditory function. Among them, FGFR1 in CDsEVs may mediate the survival of cochlear hair cells via sEVs. Finally, the joint analysis of single CDsEV sequencing and single-cell RNA sequencing data is utilized to trace cellular origins of CDsEVs. The results show that different types of cochlear cells secrete different amounts of CDsEVs, with K & ouml;lliker's organ cells and supporting cells secrete the most. The findings are expected to enhance the understanding of CDsEVs in the cochlea.

引用

页数：16

共 77 条

[1] Proteomics analysis of circulating small extracellular vesicles: Focus on the contribution of EVs to tumor metabolism [J].