水牛犊牛血浆外泌体的分离与鉴定

doi:10.16431/j.cnki.1671-7236.2021.09.010

Abstract

Abstract: The aim of this study was to separate exosomes from the plasma of buffalo calves, analysis and identify molecular biological characteristics of the isolated exosomes. The differential centrifugation was used to separate exosomes from plasma of three buffalo calves, and the morphology of extracted exosomes was observed under a transmission electron microscope. The particle size of the extracted exosomes was analyzed using nanoparticle size and Zeta potentiometer, and the expression of exosome marked proteins, including Calnexin, tumor susceptibility gene 101 (TSG101), CD9 and CD81, were detected by Western blotting method. The results showed that plasma exosomes of buffalo calves were mostly round and elliptical, with a diameter between 30 to 150 nm. The Western blotting results showed that the specific proteins Calnexin, TSG101 and CD81 were positive expressed in the extracted plasma exosomes, but protein CD9 was negative. Based on morphology and molecular characteristics, it was confirmed that the obtained extract were exosomes, indicating that the differential centrifugation could be used to successfully isolate plasma exosomes of buffalo calves. The results provided an important technical basis and reference for further research on buffalo exosome.

Key words: buffalos; plasma; exosome; differential centrifugation; exosome specific protein

CLC Number:

Q251

PANG Chunying, LIANG Shasha, WEN Chongli, CHEN Mingtang, LU Chengwei, WEI Kelong, PAN Yuhong, LIANG Xianwei. Isolation and Identification of Plasma Exosomes in Buffalo Calves[J]. China Animal Husbandry and Veterinary Medicine, 2021, 48(9): 3200-3205.

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Isolation and Identification of Plasma Exosomes in Buffalo Calves

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