Specific and Non-Invasive Fluorescent Labelling of Extracellular Vesicles for Evaluation of Intracellular Processing by Intestinal Epithelial Cells

Biomedicines
Maria S HansenJan T Rasmussen

Abstract

The presence of extracellular vesicles (EVs) in milk has gained interest due to their capacity to modulate the infant's intestinal and immune system. Studies suggest that milk EVs are enriched in immune-modulating proteins and miRNA, highlighting their possible health benefits to infants. To assess uptake of milk EVs by intestinal epithelial cells, a method was developed using labelling of isolated EVs with fluorophore-conjugated lactadherin. Lactadherin is a generic and validated EV marker, which enables an effective labelling of phosphatidylserine (PS) exposing EVs. Labelled EVs could effectively be used to describe a dose- and time-dependent uptake into the intestinal epithelial Caco-2 cell line. Additionally, fluorescence microscopy was employed to show that EVs colocalize with endosomal markers and lysosomes, indicating that EVs are taken up via general endocytotic mechanisms. Collectively, a method to specifically label isolated EVs is presented and employed to study the uptake of milk EVs by intestinal epithelial cells.

References

Feb 1, 1973·Analytical Biochemistry·G R Schacterle, R L Pollack
Mar 1, 1996·The Journal of Experimental Medicine·G RaposoH J Geuze
Sep 15, 1996·European Journal of Biochemistry·J HvarregaardT E Petersen
Jul 21, 2007·The Journal of Immunology : Official Journal of the American Association of Immunologists·Charlotte AdmyreSusanne Gabrielsson
Oct 6, 2009·The Journal of Biological Chemistry·Isabella ParoliniStefano Fais
May 4, 2010·Biochemical and Biophysical Research Communications·Taketoshi HataNaohito Aoki
Jun 10, 2010·Journal of Cellular Biochemistry·Tian TianZhongdang Xiao
Jan 18, 2011·Journal of Translational Medicine·Cecilia LässerHadi Valadi
Dec 2, 2011·Journal of Proteomics·Timothy A ReinhardtRandy E Sacco
Jun 30, 2012·Nature Methods·Johannes SchindelinAlbert Cardona
Dec 21, 2012·Journal of Cellular Physiology·Tian TianZhong-Dang Xiao
Feb 7, 2014·BMC Genomics·Ting ChenYong-Liang Zhang
Mar 13, 2014·Journal of Thrombosis and Haemostasis : JTH·N ArraudA R Brisson
Aug 22, 2014·Journal of Extracellular Vesicles·Laura Ann MulcahyDavid Raul Francisco Carter
Sep 11, 2014·Journal of Extracellular Vesicles·Marijke I ZonneveldEsther N M Nolte-'t Hoen
Mar 3, 2015·Journal of Dairy Science·Hirohisa IzumiYasuhiro Takeda
Sep 17, 2015·Early Human Development·Nicholas J AndreasKirsty Mehring Le-Doare
Nov 26, 2015·Cancer Letters·Radha MunagalaRamesh C Gupta
Mar 18, 2016·American Journal of Physiology. Cell Physiology·Rio Jati KusumaJanos Zempleni
Apr 27, 2016·The Journal of Cell Biology·Wolf HeusermannNicole C Meisner-Kober
Dec 17, 2016·Trends in Cell Biology·Sybren L N MaasAlissa M Weaver
Dec 21, 2016·Nanomedicine : Nanotechnology, Biology, and Medicine·Federica CaponnettoEnrico Ferrari
Apr 8, 2017·Journal of Extracellular Vesicles·Kristine BlansJan T Rasmussen
May 14, 2017·Scientific Reports·Aizea Morales-KastresanaJennifer C Jones
Sep 19, 2017·Journal of Controlled Release : Official Journal of the Controlled Release Society·Helena Costa VerderaPieter Vader
Mar 8, 2018·Journal of Extracellular Vesicles·Masaharu SomiyaTakahiro Ochiya
Jun 16, 2018·Journal of Extracellular Vesicles·Abderrahim BenmoussaPatrick Provost
Apr 6, 2019·Molecular Pharmaceutics·Jessica E PullanSanku Mallik
Feb 26, 2020·Animals : an Open Access Journal From MDPI·Suyu QuanBenhai Xiong
Mar 15, 2020·Nutrients·Jeffrey D Galley, Gail E Besner

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Citations

Aug 28, 2021·Journal of Controlled Release : Official Journal of the Controlled Release Society·Anisha D'SouzaDevika S Manickam

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Methods Mentioned

BETA
size exclusion chromatography
light scattering
confocal microscopy
Assay
transfection
fluorescence microscopy
GTPases
IEC-6

Software Mentioned

Fiji
GraphPad
ImageJ
GraphPad Prism

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