33
4 Notes
1. The ultrasonic probe should be washed with H 2 SO 4 , ultrapure
water and ethanol, respectively.
2. The ultrasonication step should be carried out on ice in order to
prevent heating of the tissues due to high-frequency
sonication.
3. Uranyl acetate solution should be filtered from 0.22 μm poresized syringe filter in order to prevent accumulation of large or
aggregated particles onto the grid.
4. A260/280 ratio for RNA quantification should be around 2,
indicating the purity of RNA. If this ratio is over/lower, a
protein- caused impurity is a question that requires an extensive
purification step.
Acknowledgments
This work has been co-financed by the European Union and Greek
national funds through the program “Support for Researchers
with Emphasis on Young Researchers” (call code: EDBM34, ΚΕ
14995) and under the research title “Preparation and study of
innovative forms of administration of pharmaceutical molecules
targeting at improved pharmacological properties.”
References
1. Melo SA, Luecke LB, Kahlert C et al (2015)
Glypican-1 identifies cancer exosomes and
detects early pancreatic cancer. Nature
523:177–182
2. Malla B, Zaugg K, Vassella E et al (2017)
Exosomes and exosomal microRNAs in prostate cancer radiation therapy. Int J Radiat
Oncol Biol Phys 98(5):982–995
3. Agrawal AK, Aqil F, Jeyabalan J et al (2017)
Milk-derived exosomes for oral delivery of
paclitaxel. Nanomedicine 13(5):1627–1636
4. Chen B, Li Q, Zhao B et al (2017) Stem cellderived extracellular vesicles as a novel potential therapeutic tool for tissue repair. Stem
Cells Transl Med 6:1753–1758. https://doi.
org/10.1002/sctm.16-0477
5. Leblanc P, Arellano-Anaya ZE, Bernard E et al
(2017) Isolation of exosomes and microvesicles
from cell culture systems to study prion transmission. Methods Mol Biol 1545:153–176
6. Garcia-Contreras M, Shah SH, Tamayo A
et al (2017) Plasma-derived exosome characterization reveals a distinct microRNA signature in long duration type 1 diabetes. Sci Rep
7(1):5998
7. Helwa I, Cai J, Drewry MD et al (2017) A
comparative study of serum exosome isolation using differential ultracentrifugation
and three commercial reagents. PLoS One
12(1):e0170628
8. Miranda KC, Bond DT, Levin JZ et al (2014)
Massively parallel sequencing of human urinary exosome/microvesicle RNA reveals a
predominance of non-coding RNA. PLoS One
9(5):e96094
9. Vojtech L, Woo S, Hughes S et al (2014)
Exosomes in human semen carry a distinctive
repertoire of small non-coding RNAs with
potential regulatory functions. Nucleic Acids
Res 42(11):7290–7304
10. Perkins EJ, Bao W, Guan X et al (2006)
Comparison of transcriptional responses in
liver tissue and primary hepatocyte cell cultures
after exposure to hexahydro-1, 3, 5-trinitro-1,
Ultrasonics-Assisted Effective Isolation and Characterization of Exosomes from Whole…
4 Notes
1. The ultrasonic probe should be washed with H 2 SO 4 , ultrapure
water and ethanol, respectively.
2. The ultrasonication step should be carried out on ice in order to
prevent heating of the tissues due to high-frequency
sonication.
3. Uranyl acetate solution should be filtered from 0.22 μm poresized syringe filter in order to prevent accumulation of large or
aggregated particles onto the grid.
4. A260/280 ratio for RNA quantification should be around 2,
indicating the purity of RNA. If this ratio is over/lower, a
protein- caused impurity is a question that requires an extensive
purification step.
Acknowledgments
This work has been co-financed by the European Union and Greek
national funds through the program “Support for Researchers
with Emphasis on Young Researchers” (call code: EDBM34, ΚΕ
14995) and under the research title “Preparation and study of
innovative forms of administration of pharmaceutical molecules
targeting at improved pharmacological properties.”
References
1. Melo SA, Luecke LB, Kahlert C et al (2015)
Glypican-1 identifies cancer exosomes and
detects early pancreatic cancer. Nature
523:177–182
2. Malla B, Zaugg K, Vassella E et al (2017)
Exosomes and exosomal microRNAs in prostate cancer radiation therapy. Int J Radiat
Oncol Biol Phys 98(5):982–995
3. Agrawal AK, Aqil F, Jeyabalan J et al (2017)
Milk-derived exosomes for oral delivery of
paclitaxel. Nanomedicine 13(5):1627–1636
4. Chen B, Li Q, Zhao B et al (2017) Stem cellderived extracellular vesicles as a novel potential therapeutic tool for tissue repair. Stem
Cells Transl Med 6:1753–1758. https://doi.
org/10.1002/sctm.16-0477
5. Leblanc P, Arellano-Anaya ZE, Bernard E et al
(2017) Isolation of exosomes and microvesicles
from cell culture systems to study prion transmission. Methods Mol Biol 1545:153–176
6. Garcia-Contreras M, Shah SH, Tamayo A
et al (2017) Plasma-derived exosome characterization reveals a distinct microRNA signature in long duration type 1 diabetes. Sci Rep
7(1):5998
7. Helwa I, Cai J, Drewry MD et al (2017) A
comparative study of serum exosome isolation using differential ultracentrifugation
and three commercial reagents. PLoS One
12(1):e0170628
8. Miranda KC, Bond DT, Levin JZ et al (2014)
Massively parallel sequencing of human urinary exosome/microvesicle RNA reveals a
predominance of non-coding RNA. PLoS One
9(5):e96094
9. Vojtech L, Woo S, Hughes S et al (2014)
Exosomes in human semen carry a distinctive
repertoire of small non-coding RNAs with
potential regulatory functions. Nucleic Acids
Res 42(11):7290–7304
10. Perkins EJ, Bao W, Guan X et al (2006)
Comparison of transcriptional responses in
liver tissue and primary hepatocyte cell cultures
after exposure to hexahydro-1, 3, 5-trinitro-1,
Ultrasonics-Assisted Effective Isolation and Characterization of Exosomes from Whole…