25
Thomas Mavromoustakos et al. (eds.), Supramolecules in Drug Discovery and Drug Delivery: Methods and Protocols,
Methods in Molecular Biology, vol. 2207, https://doi.org/10.1007/978-1-0716-0920-0_3,
© Springer Science+Business Media, LLC, part of Springer Nature 2021
Chapter 3
Ultrasonics-Assisted Effective Isolation
and Characterization of Exosomes from Whole Organs
Burak Derkus and Emel Emregul
Abstract
Exosomes, natural and nanovesicular structures surrounded by a lipid membrane, tend to be secreted
toward extracellular environments by almost all cell types. Late studies have shown them to be effective in
several complex biological processes like cancer development and metastasis, immune system regulation,
cellular signal transduction, stem cell differentiation, and regeneration of damaged tissues. Although there
are many studies dealing with the role of exosomes in the aforementioned fields, the mechanisms remained
largely unknown. There is therefore a need for further study on exosome isolation from different sources.
While researchers mostly use serum, plasma, urine, and cell culture media as a source for exosome isolation, there are no studies dealing with direct isolation of exosomes from whole organs in literature. In this
study, we propose a protocol for effective isolation of exosomes from whole organs. Mouse brain, heart,
and liver were chosen as the sources of exosomes in this study. Isolated exosomes were successfully characterized with BCA test, western blot, transmission electron microscopy and ELISA.
Key words Extracellular vesicles, Nanovesicles, Exosomes, Isolation, Ultrasonication
1 Introduction
Exosomes, which are released by almost all kinds of cells to the
extracellular environment, are lipid-involved natural nanovesicles
with a diameter of 50–150 nm. Although they were evaluated as
cellular wastes in the early studies, their potential of use in diagnosis and treatment of diseases was recognized after 2000s. Owing to
their rich microRNA (miRNA) content, which is 60 times higher
than regular biological fluids, exosomes today are utilized in diagnostics [1], cancer treatment [2], drug delivery [3] and stem cellbased technologies [4]. Exosomes are mainly isolated from cell
culture wastes [5], plasma [6], serum [7], urine [8], and semen
[9]. While they are mostly and practically isolated from cell cultureconditioned media (CM), it is thought that exosomal content
might be unsteady due to genotypic and phenotypic differences
Thomas Mavromoustakos et al. (eds.), Supramolecules in Drug Discovery and Drug Delivery: Methods and Protocols,
Methods in Molecular Biology, vol. 2207, https://doi.org/10.1007/978-1-0716-0920-0_3,
© Springer Science+Business Media, LLC, part of Springer Nature 2021
Chapter 3
Ultrasonics-Assisted Effective Isolation
and Characterization of Exosomes from Whole Organs
Burak Derkus and Emel Emregul
Abstract
Exosomes, natural and nanovesicular structures surrounded by a lipid membrane, tend to be secreted
toward extracellular environments by almost all cell types. Late studies have shown them to be effective in
several complex biological processes like cancer development and metastasis, immune system regulation,
cellular signal transduction, stem cell differentiation, and regeneration of damaged tissues. Although there
are many studies dealing with the role of exosomes in the aforementioned fields, the mechanisms remained
largely unknown. There is therefore a need for further study on exosome isolation from different sources.
While researchers mostly use serum, plasma, urine, and cell culture media as a source for exosome isolation, there are no studies dealing with direct isolation of exosomes from whole organs in literature. In this
study, we propose a protocol for effective isolation of exosomes from whole organs. Mouse brain, heart,
and liver were chosen as the sources of exosomes in this study. Isolated exosomes were successfully characterized with BCA test, western blot, transmission electron microscopy and ELISA.
Key words Extracellular vesicles, Nanovesicles, Exosomes, Isolation, Ultrasonication
1 Introduction
Exosomes, which are released by almost all kinds of cells to the
extracellular environment, are lipid-involved natural nanovesicles
with a diameter of 50–150 nm. Although they were evaluated as
cellular wastes in the early studies, their potential of use in diagnosis and treatment of diseases was recognized after 2000s. Owing to
their rich microRNA (miRNA) content, which is 60 times higher
than regular biological fluids, exosomes today are utilized in diagnostics [1], cancer treatment [2], drug delivery [3] and stem cellbased technologies [4]. Exosomes are mainly isolated from cell
culture wastes [5], plasma [6], serum [7], urine [8], and semen
[9]. While they are mostly and practically isolated from cell cultureconditioned media (CM), it is thought that exosomal content
might be unsteady due to genotypic and phenotypic differences