Lyophilization Based Isolation of Exosomes

Author:

Qazi Rida e Maria1,Sajid Zahra1,Zhao Chunqiu2,Hussain Irfan1ORCID,Iftikhar Fizza1,Jameel Muhammad1,Rehman Fawad Ur1ORCID,Mian Afsar Ali1

Affiliation:

1. Centre for Regenerative Medicine and Stem Cells Research, The Aga Khan University, 1st Flour, Juma Building, Stadium Road, Karachi 74800, Sindh, Pakistan

2. State Key Lab of Bioelectronics, Southeast University, Sipailou 2, Nanjing 210096, China

Abstract

Exosomes are nanoscale extracellular vesicles which regulate intercellular communication. They have great potential for application in nanomedicine. However, techniques for their isolation are limited by requirements for advanced instruments and costly reagents. In this study, we developed a lyophilization-based method for isolating exosomes from cultured cells. The isolated exosomes were characterized for protein content using Bradford assay, and for size distribution and shape using scanning electron microscopy (SEM) and nanoparticles tracking analysis (NTA). In addition, CD63, CD9, CD81, HSP70 and TSG101 were evaluated as essential exosomal surface markers using Western blot. Drug loading and release studies were performed to confirm their drug delivery properties using an in vitro model. Exosomes were also loaded with commercial dyes (Cy5, Eosin) for the evaluation of their drug delivery properties. All these characterizations confirmed successful exosome isolation with measurements of less than 150 nm, having a typical shape, and by expressing the known exosome surface protein markers. Finally, tyrosine kinase inhibitors (dasatinib and ponatinib) were loaded on the exosomes to evaluate their anticancer effects on leukemia cells (K562 and engineered Ba/F3-BCR-ABL) using MTT and Annexin-PI assays. The expression of MUC1 protein on the exosomes isolated from MCF-7 cells also indicated that their potential diagnostic properties were intact. In conclusion, we developed a new method for exosome isolation from cultured cells. These exosomes met all the essential requirements in terms of characterization, drug loading and release ability, and inhibition of proliferation and apoptosis induction in Ph+ leukemia cells. Based on these results, we are confident in presenting the lyophilization-based exosome isolation method as an alternative to traditional techniques for exosome isolation from cultured cells.

Funder

Higher Education Commission

Publisher

MDPI AG

Subject

Inorganic Chemistry,Organic Chemistry,Physical and Theoretical Chemistry,Computer Science Applications,Spectroscopy,Molecular Biology,General Medicine,Catalysis

Reference49 articles.

1. Nanotechnology-based promising strategies for the management of COVID-19: Current development and constraints;Rai;Expert Rev. Anti-Infect. Ther.,2022

2. Nanomedicine for COVID-19: The role of nanotechnology in the treatment and diagnosis of COVID-19;Vahedifard;Emerg. Mater.,2021

3. Metallic nanoscale-knife application in cancer theranostics;Zhao;Smart Mater. Med.,2023

4. Nanomedicine: Why it still taking long from “bench to bedside”;Rehman;Biomed. Lett.,2018

5. Toxicological assessment of nanoparticles and microplastics;Younas;Biomed. Lett.,2022

同舟云学术

1.学者识别学者识别

2.学术分析学术分析

3.人才评估人才评估

"同舟云学术"是以全球学者为主线,采集、加工和组织学术论文而形成的新型学术文献查询和分析系统,可以对全球学者进行文献检索和人才价值评估。用户可以通过关注某些学科领域的顶尖人物而持续追踪该领域的学科进展和研究前沿。经过近期的数据扩容,当前同舟云学术共收录了国内外主流学术期刊6万余种,收集的期刊论文及会议论文总量共计约1.5亿篇,并以每天添加12000余篇中外论文的速度递增。我们也可以为用户提供个性化、定制化的学者数据。欢迎来电咨询!咨询电话:010-8811{复制后删除}0370

www.globalauthorid.com

TOP

Copyright © 2019-2024 北京同舟云网络信息技术有限公司
京公网安备11010802033243号  京ICP备18003416号-3