TY - JOUR
T1 - Label-free characterization and real-time monitoring of cell uptake of extracellular vesicles
AU - Koponen, Annika
AU - Kerkelä, Erja
AU - Rojalin, Tatu
AU - Lázaro-Ibáñez, Elisa
AU - Suutari, Teemu
AU - Saari, Heikki O.
AU - Siljander, Pia
AU - Yliperttula, Marjo
AU - Laitinen, Saara
AU - Viitala, Tapani
N1 - Funding Information:
The authors thank Viikki Flow Cytometry Core Facility, EV Core Facility and the Light Microscopy Unit, Institute of Biotechnology (all University of Helsinki). Lotta Sankkila, Lotta Andersson and Birgitta Rantala are acknowledged for their excellent technical assistance. Funding: Finnish Funding Agency for Technology/Business Finland (Grant nos. 440211 (AK, MY), 440935 (PS, MY, TV), Salwe GID (SL)), Sigrid Juselius Foundation (TR), Doctoral Programme in Materials Research and Nanoscience (TS), Alfred Kordelin Foundation (EL-I), Emil Aaltonen foundation (HOS), Academy of Finland (Grant nos. 287089 (PS), 294309 (TV, AK)) and Magnus Ehrnrooth Foundation (PS) are gratefully acknowledged.
Funding Information:
The authors thank Viikki Flow Cytometry Core Facility, EV Core Facility and the Light Microscopy Unit, Institute of Biotechnology (all University of Helsinki). Lotta Sankkila, Lotta Andersson and Birgitta Rantala are acknowledged for their excellent technical assistance. Funding: Finnish Funding Agency for Technology/Business Finland (Grant nos. 440211 (AK, MY), 440935 (PS, MY, TV), Salwe GID (SL)), Sigrid Juselius Foundation (TR), Doctoral Programme in Materials Research and Nanoscience (TS), Alfred Kordelin Foundation (EL-I), Emil Aaltonen foundation (HOS), Academy of Finland (Grant nos. 287089 (PS), 294309 (TV, AK)) and Magnus Ehrnrooth Foundation (PS) are gratefully acknowledged.
Publisher Copyright:
© 2020 Elsevier B.V.
PY - 2020/11/15
Y1 - 2020/11/15
N2 - Extracellular vesicles (EVs) have the ability to function as molecular vehicles and could therefore be harnessed to deliver drugs to target cells in diseases such as cancer. The composition of EVs determines their function as well as their interactions with cells, which consequently affects the cell uptake efficacy of EVs. In this study, we present two novel label-free approaches for studying EVs; characterization of EV composition by time-gated surface-enhanced Raman spectroscopy (TG-SERS) and monitoring the kinetics and amount of cellular uptake of EVs by surface plasmon resonance (SPR) in real-time. Using these methods, we characterized the most abundant EVs of human blood, red blood cell (RBC)- and platelet (PLT)-derived EVs and studied their interactions with prostate cancer cells. Complementary studies were performed with nanoparticle tracking analysis for concentration and size determinations of EVs, zeta potential measurements for surface charge analysis, and fluorophore-based confocal imaging and flow cytometry to confirm EV uptake. Our results revealed distinct biochemical features between the studied EVs and demonstrated that PLT-derived EVs were more efficiently internalized by PC-3 cells than RBC-derived EVs. The two novel label-free techniques introduced in this study were found to efficiently complement conventional techniques and paves the way for further use of TG-SERS and SPR in EV studies.
AB - Extracellular vesicles (EVs) have the ability to function as molecular vehicles and could therefore be harnessed to deliver drugs to target cells in diseases such as cancer. The composition of EVs determines their function as well as their interactions with cells, which consequently affects the cell uptake efficacy of EVs. In this study, we present two novel label-free approaches for studying EVs; characterization of EV composition by time-gated surface-enhanced Raman spectroscopy (TG-SERS) and monitoring the kinetics and amount of cellular uptake of EVs by surface plasmon resonance (SPR) in real-time. Using these methods, we characterized the most abundant EVs of human blood, red blood cell (RBC)- and platelet (PLT)-derived EVs and studied their interactions with prostate cancer cells. Complementary studies were performed with nanoparticle tracking analysis for concentration and size determinations of EVs, zeta potential measurements for surface charge analysis, and fluorophore-based confocal imaging and flow cytometry to confirm EV uptake. Our results revealed distinct biochemical features between the studied EVs and demonstrated that PLT-derived EVs were more efficiently internalized by PC-3 cells than RBC-derived EVs. The two novel label-free techniques introduced in this study were found to efficiently complement conventional techniques and paves the way for further use of TG-SERS and SPR in EV studies.
KW - Cell uptake
KW - Extracellular vesicles
KW - Prostate cancer cells
KW - Surface plasmon resonance
KW - Time-gated surface-enhanced Raman spectroscopy
UR - http://www.scopus.com/inward/record.url?scp=85089887653&partnerID=8YFLogxK
U2 - 10.1016/j.bios.2020.112510
DO - 10.1016/j.bios.2020.112510
M3 - Article
C2 - 32877783
AN - SCOPUS:85089887653
SN - 0956-5663
VL - 168
JO - Biosensors and Bioelectronics
JF - Biosensors and Bioelectronics
M1 - 112510
ER -