双峰驼血浆外泌体分离提取及肝癌细胞对外泌体摄取的实验研究
作者:
作者单位:

1.内蒙古农业大学兽医学院 呼和浩特 010018;2.农业农村部动物疾病临床诊疗技术重点实验室 呼和浩特 010018

作者简介:

于雯,女,硕士研究生;研究方向:动物生理和病理研究;E-mail:2219245084@qq.com。

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中图分类号:

Q955

基金项目:

国家自然科学基金项目(No. 32160821,31860693),内蒙古自治区自然科学基金项目(No. 2023MS03021);


Isolation and Extraction of Plasma Exosomes From Bactrian Camels and Experimental Study on Exosome Uptake by Hepatocellular Carcinoma Cells
Author:
Affiliation:

1.College of Veterinary Medicine, Inner Mongolia Agricultural University, Hohhot 010018; 2.Key Laboratory of Clinical Diagnosis and Treatment of Animal Diseases, Ministry of Agriculture, Hohhot 010018, China

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    摘要:

    为进一步研究双峰驼(Camelus bactrianus)血浆来源的外泌体在肝癌细胞侵袭和转移中的调节作用,本研究采用超速离心法分离双峰驼血浆外泌体,并通过透射电镜观察其形态和超微结构。随后,使用纳米粒子跟踪分析仪测定外泌体的粒径及分布。将荧光染料PKH67标记的外泌体与肝癌细胞(MHCC-97H和Huh7细胞)共培养,并应用扫描共聚焦显微镜(LSCM)观察肝癌细胞对双峰驼血浆来源外泌体的摄取情况。结果显示,分离获得的双峰驼血浆外泌体均呈现杯状或圆形形态,直径约为100 nm;纳米粒子跟踪分析的结果表明,其颗粒大小的主峰分别为约124.6 nm和117.9 nm,总体尺寸分布范围从50 nm到200 nm。在扫描共聚焦显微镜下观察,PKH67标记的外泌体能够进入MHCC-97H和Huh7肝癌细胞内,且主要分布于细胞膜及核周的细胞质中。本研究成功提取了双峰驼血浆外泌体,并观察到肝癌细胞对外泌体的摄取现象,表明肝癌细胞可能是双峰驼外泌体的靶细胞之一。

    Abstract:

    [Objectives] This study aims to study the regulatory role of exosomes derived from Bactrian Camel Camelus bactrianus plasma in the invasion and metastasis of hepatocellular carcinoma cells. [Methods] In this study, the plasma exosomes of Bactrian Camels were separated by ultracentrifugation, the morphology and ultrastructure of the exosomes were observed by transmission electron microscopy, and the particle size and distribution of the exosomes were determined by nanoparticle tracking analyzer, the exosomes were co-cultured with hepatoma cells (MHCC-97H and Huh7 cells) after labeling with fluorescent dye PKH67, and the uptake of plasma-derived exosomes by hepatoma cells was observed by scanning confocal microscopy (LSCM). [Results] The obtained plasma exosomes of Bactrian Camels were all cup-shaped or round-shaped, with a diameter of about 100 nm (Fig. 1):the main peaks of particle size were about 124.6 nm and 117.9 nm, respectively, and the overall size distribution ranged from 50 nm to 200 nm (Fig. 2). Exosomes carrying PKH67 markers were observed to enter MHCC-97H and Huh7 cells under LSCM, and mainly distributed in the cytoplasm of cell membranes and perinucleus (Fig. 3). [Conclusion] In this study, exosomes derived from Bactrian Camel plasma were successfully extracted, and the uptake of exosomes by hepatoma cells was observed, suggesting that hepatocellular carcinoma cells may be one of the target cells of Bactrian exosomes.

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引用本文

于雯,李子怡,牛雅如,贺星川,白浩宇,玉斯日古楞.2025.双峰驼血浆外泌体分离提取及肝癌细胞对外泌体摄取的实验研究.动物学杂志,60(2):239-246.

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  • 收稿日期:2024-05-21
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  • 在线发布日期: 2025-04-25
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