Isolation, identification, and miRNAs enrichment analysis of exosomes derived from giant panda umbilical cord mesenchymal stem cells

AN Junhui; LI Yuan; WANG Donghui; CHEN Jiasong; LI Hongyan; LIANG Xiaohu; FENG Tongying; CAI Zhigang; HOU Rong; ZENG Changjun; LIU Yuliang

doi:10.16829/j.slxb.150664

2023 , Vol. 43 >Issue 1: 33 - 40

DOI: https://doi.org/10.16829/j.slxb.150664

ORIGINAL PAPERS

Isolation, identification, and miRNAs enrichment analysis of exosomes derived from giant panda umbilical cord mesenchymal stem cells

AN Junhui ,
LI Yuan ,
WANG Donghui ,
CHEN Jiasong ,
LI Hongyan ,
LIANG Xiaohu ,
FENG Tongying ,
CAI Zhigang ,
HOU Rong ,
ZENG Changjun ,
LIU Yuliang

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1 College of Animal Sciences and Technology, and Farm Animal Genetic Resources Exploration and Innovation Key Laboratory of Si-chuan Province, Sichuan Agricultural University, Chengdu 611130, China;
2 Chengdu Research Base of Giant Panda Breeding, Sichuan Key Laboratory of Conservation Biology for Endangered Wildlife, Cheng-du 610081, China;
3 Sichuan Academy of Giant Panda, Chengdu 610081, China

Received date: 2022-02-25

Revised date: 2022-07-05

Online published: 2023-01-10

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Abstract

The exosomes released by giant panda umbilical cord mesenchymal stem cells (UC-MSCs) through paracrine mechanisms have application prospects in giant panda veterinary care and disease treatment. The aims of this study were to explore a method of separating exosomes secreted by giant panda UC-MSCs, identify their biological and molecular characteristics, and study the miRNA expression profile and distribution characteristics. Ultracentrifugation was used to successfully isolate exosomes from the supernatant of giant panda UC-MSCs culture medium. The morphological observation was performed by transmission electron microscopy, particle size was detected by nanoparticle tracking analyzer, and the expression of specific molecular markers was detected by western blotting. In addition, the small RNA sequencing technology was used to explore the expression profile and function of exosomal miRNAs. The results showed that the exosomes obtained from giant panda UC-MSCs displayed a circular cup-shaped structure with a particle size of (79. 15 ± 4. 81) nm. These exosomes were positive for CD81 and TSG101 but negative for CALNEXIN. The miRNAs in exosomes were mainly composed of miR-148-3p (30. 28%) and miR-21-5p (21. 72%). In this study, we harvested exosomes from the supernatant of giant panda UC-MSCs culture medium for the first time and conducted the enrichment analysis and functional prediction of UC-MSCs miRNAs. This study lays the foundation for follow-up studies of UCMSCs exosomes and their clinical application in giant pandas.

Key words： Giant panda; Umbilical cord mesenchymal stem cells; Exosomes; miRNA

Cite this article

AN Junhui , LI Yuan , WANG Donghui , CHEN Jiasong , LI Hongyan , LIANG Xiaohu , FENG Tongying , CAI Zhigang , HOU Rong , ZENG Changjun , LIU Yuliang . Isolation, identification, and miRNAs enrichment analysis of exosomes derived from giant panda umbilical cord mesenchymal stem cells[J]. ACTA THERIOLOGICA SINICA, 2023 , 43(1) : 33 -40 . DOI: 10.16829/j.slxb.150664

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