ACS Applied Bio Materials,
Journal Year:
2024,
Volume and Issue:
7(11), P. 7770 - 7783
Published: Nov. 1, 2024
Extracellular
vesicles
(EVs)
are
cell-secreted
lipid
bilayer
delimited
particles
that
mediate
cellular
communication.
These
tiny
sacs
of
information
play
an
important
role
in
cell
communication
and
alter
the
physiological
process
under
both
normal
pathological
conditions.
As
such,
tracking
EVs
can
provide
valuable
regarding
basic
understanding
communication,
onset
early
malignancy,
biomarker
discovery.
Most
current
EV-tracking
strategies
invasive,
altering
natural
characteristics
by
modifying
with
lipophilic
dyes
or
surface
proteins
fluorescent
reporters.
The
invasive
labeling
could
processes
thereby
have
major
limitations
for
functional
studies.
Here,
we
report
alternative
minimally
EV
strategy
using
PicoGreen
(PG),
a
small
molecule
fluoresces
at
520
nm
when
bound
to
dsDNA.
We
show
PG
binds
dsDNA
associated
(50–200
nm),
forming
stable
highly
PG-DNA
complex
(PG-EVs).
In
2D
culture
3D
organoid
models,
PG-EV
showed
efficient
properties,
including
high
signal-to-noise
ratio,
time-
concentration-dependent
uptake,
ability
traverse
environment.
further
validated
dual-labeled
following
two
orthogonal
strategies:
(1)
Bioconjugation
via
amine
(2)
donor
engineering
endogenously
expressing
mCherry-tetraspanin
(CD9/CD63/CD81)
reporter
proteins.
Our
study
has
shown
feasibility
as
effective
be
applied
studying
across
multiple
model
systems.
Advanced Science,
Journal Year:
2024,
Volume and Issue:
11(30)
Published: June 14, 2024
Abstract
In
recent
decades,
research
on
Extracellular
Vesicles
(EVs)
has
gained
prominence
in
the
life
sciences
due
to
their
critical
roles
both
health
and
disease
states,
offering
promising
applications
diagnosis,
drug
delivery,
therapy.
However,
inherent
heterogeneity
complex
origins
pose
significant
challenges
preparation,
analysis,
subsequent
clinical
application.
This
review
is
structured
provide
an
overview
of
biogenesis,
composition,
various
sources
EVs,
thereby
laying
groundwork
for
a
detailed
discussion
contemporary
techniques
preparation
analysis.
Particular
focus
given
state‐of‐the‐art
technologies
that
employ
microfluidic
non‐microfluidic
platforms
EV
processing.
Furthermore,
this
discourse
extends
into
innovative
approaches
incorporate
artificial
intelligence
cutting‐edge
electrochemical
sensors,
with
particular
emphasis
single
proposes
current
outlines
prospective
avenues
future
research.
The
objective
motivate
researchers
innovate
expand
methods
analysis
fully
unlocking
biomedical
potential.
Frontiers in Immunology,
Journal Year:
2024,
Volume and Issue:
15
Published: July 8, 2024
Exosome-derived
microRNAs
(miRNAs)
are
biomacromolecules
and
nanoscale
extracellular
vesicles
originating
from
intracellular
compartments
that
secreted
by
most
cells
into
the
space.
This
review
examines
formation
function
of
exosomal
miRNAs
in
biological
information
transfer,
explores
pathogenesis
vitiligo,
highlights
relationship
between
vitiligo.
The
aim
is
to
deepen
understanding
how
influence
immune
imbalance,
oxidative
stress
damage,
melanocyte-keratinocyte
interactions,
melanogenesis
disorders
development
enhanced
may
contribute
potential
diagnostic
therapeutic
options
for
Journal of Biological Chemistry,
Journal Year:
2025,
Volume and Issue:
unknown, P. 108481 - 108481
Published: April 1, 2025
Cells
across
biological
kingdoms
release
extracellular
vesicles
(EVs)
as
a
means
of
communication
with
other
cells,
be
their
friends
or
foes.
This
is
indeed
true
for
the
intracellular
malaria
parasite
Plasmodium
falciparum
(Pf),
which
utilizes
EVs
to
transport
bioactive
molecules
various
human
host
systems.
Yet,
study
this
mode
in
research
currently
constrained
due
limitations
high-resolution
tools
and
absence
commercial
antibodies.
Here,
we
demonstrate
power
an
advanced
spectral
flow
cytometry
approach
robustly
detect
secreted
EVs,
isolated
from
Pf-infected
red
blood
cells.
By
labeling
both
EV
membrane
lipids
DNA
cargo
within
(non-antibody
staining
approach),
were
able
subpopulation
parasitic-derived
enriched
DNA.
Furthermore,
could
quantitatively
measure
DNA-carrying
two
distinct
stages
parasite:
rings
trophozoites.
Our
findings
showcase
potential
monitor
dynamic
changes
nucleic
acid
pathogenic
EVs.
PLoS ONE,
Journal Year:
2025,
Volume and Issue:
20(1), P. e0317689 - e0317689
Published: Jan. 24, 2025
Extracellular
vesicles
(EVs)
are
membrane-bound
structures
produced
and
released
into
the
extracellular
space
by
all
types
of
cells.
Due
to
their
characteristics,
EVs
play
crucial
roles
in
cellular
communication
signaling,
holding
an
immense
potential
as
biomarkers
molecular
transporters.
Various
methods
have
been
developed
label
characterize
EVs,
however,
visualizing
remains
a
process
that
requires
highly
specialized
expensive
equipment,
which
is
not
always
available
laboratories.
In
this
study,
we
adapted
protocol
originally
designed
for
analysis
flow
cytometry
using
Calcein-AM,
convert
it
useful
effective
tool
epifluorescence
microscopy
coupled
with
deconvolution
system.
This
approach
can
be
very
basic
analyses,
enabling
researchers
verify
distribution
internalization
across
Such
insights
guide
decisions
on
whether
advance
more
detailed
confocal
or
perform
additional
assays.
Small Methods,
Journal Year:
2025,
Volume and Issue:
unknown
Published: Feb. 25, 2025
Abstract
Milk‐derived
extracellular
vesicles
(mEVs)
are
promising
therapeutic
delivery
platforms
due
to
their
natural
bioactivity,
biocompatibility,
and
ability
cross
biological
barriers.
However,
analyzing
cellular
uptake
trafficking
is
limited
by
existing
fluorescent
labeling
methods,
which
often
cause
dye
leakage
disrupt
vesicle
integrity.
Here,
a
glycan‐anchored
fluorescence
strategy
for
mEVs
developed,
involving
periodate
oxidation
of
surface
sialic
acids
followed
aniline‐catalyzed
ligation
hydrazide‐functionalized
fluorophores.
Nano‐flow
cytometry
characterization
confirmed
≈100%
efficiency
without
compromising
integrity
or
behavior.
This
approach
enabled
quantitative
analysis
internalization,
identifying
clathrin‐mediated
endocytosis
macropinocytosis
as
the
primary
pathways
confirming
mEVs’
capacity
lysosomal
escape.
Comparative
analyses
showed
that
traditional
lipophilic
dyes
induced
aggregation,
leakage,
transfer,
potentially
misrepresenting
Additionally,
co‐labeling
with
fluorophores
FITC‐conjugated
paclitaxel
real‐time
tracking
drug
delivery,
revealing
burst
release
from
lysosomes
led
significant
cytotoxicity.
Overall,
allows
precise
intracellular
fate,
paving
way
further
research
application
in
targeted
delivery.
Frontiers in Bioengineering and Biotechnology,
Journal Year:
2024,
Volume and Issue:
12
Published: Sept. 18, 2024
A
well-designed
fluorescence-based
analysis
of
extracellular
vesicles
(EV)
can
provide
insights
into
the
size,
morphology,
and
biological
function
EVs,
which
be
used
in
medical
applications.
Fluorescent
nanoparticle
tracking
with
appropriate
controls
reliable
data
for
size
concentration
measurements,
while
nanoscale
flow
cytometry
is
most
tool
characterizing
molecular
cargoes.
Label
selection
a
crucial
element
all
fluorescence
methods.
The
comprehensive
obtained
if
several
labeling
approaches
given
marker
are
used,
as
they
would
complementary
information
about
EV
populations
interactions
cells.
In
EV-related
experiments,
influence
lipoproteins
protein
corona
on
results
should
considered.
By
reviewing
considering
factors
affecting
methods
techniques,
we
assert
that
will
accurate
possible
true
biology
offer
precise,
clinically
applicable
future
EV-based
diagnostic
or
therapeutic
ACS Applied Nano Materials,
Journal Year:
2024,
Volume and Issue:
7(20), P. 24160 - 24171
Published: Oct. 9, 2024
Stroke
is
a
leading
cause
of
death
and
disability
worldwide.
Extracellular
vesicles
(EVs)
derived
from
human
mesenchymal
stem
cells
(hMSCs)
offer
unique
promising
alternative
to
direct
cell
injection
as
part
cell-based
therapy
for
stroke
treatment.
The
development
labeling
strategies
essential
identifying
the
initial
biodistribution
clearance
EV-based
therapeutics.
In
this
study,
hMSC-EVs
were
labeled
with
ultrasmall
superparamagnetic
iron
oxide
(USPIO)
nanoparticles
magnetic
resonance
imaging
(MRI).
Two
methods
preparation
evaluated
after
EVs
sonicated
in
presence
USPIO
nanoparticles.
purified
by
(1)
ultracentrifugation
only
or
(2)
an
extension
harvesting
approach
that
employs
poly(ethylene
glycol)
(PEG)
enrich
EVs.
Following
vitro
assessment,
applied
ischemic
model
imaged
both
immediately
longitudinally
using
MRI.
assessment
showed
EV
characteristics
nanoparticle
labeling.
PEG
method
exhibited
3.6-fold
enhancement
contrast
equivalent
concentration
0.5
mg/mL
acquisition
parameters
(TE
=
3.5
ms,
TR
5
s)
when
dilution
factor
considered.
Sufficient
was
achieved
visualize
assess
therapeutic
potential.
Taken
together,
simultaneous
enrichment
enhanced
MRI
improved
outcomes
respect
delivery
recovery.
ACS Applied Bio Materials,
Journal Year:
2024,
Volume and Issue:
unknown
Published: Dec. 5, 2024
Exosomes
are
small
extracellular
vesicles
(EVs)
constituting
fully
biological,
cell-derived
nanovesicles
with
great
potential
in
cell-to-cell
communication
and
drug
delivery
applications.
The
current
gold
standard
for
EV
labeling
tracking
is
represented
by
fluorescent
lipophilic
dyes
which,
however,
importantly
lack
selectivity,
due
to
their
unconditional
affinity
lipids.
Herein,
an
alternative
approach
in-depth
evaluated,
taking
advantage
of
green
protein
(GFP)
farnesylation
(GFP-f),
a
post-translational
modification
directly
anchor
GFP
the
membrane.
performance
GFP-f
analyzed,
terms
selectivity
efficiency,
several
typical
experimental
setups
such
as
recipient
cells,
surface
engineering,
cargo
loading.
First,
capability
label
exosomes
was
compared,
showing
significantly
higher
levels
fluorescence
intensity
GFP-f-
than
GFP-labeled
exosomes,
highlighting
anchoring
cell
Then,
tag
further
compared
Vybrant
DiD
dye
exosome
uptake
studies,
capturing
intracellular
time-
concentration-dependent
manner.
internalization
assay
revealed
particular
ability
monitor
tagged
into
significant
peak
reached
12
h
after
administration
but
not
Vybrant-labeled
EVs.
Finally,
challenged
presence
transfection
siRNA
Results
showed
that
both
procedures
can
influence
naïve
although
maintained
cases.
Overall,
these
data
provide
direct
insight
advantages
limitations
tagging
selectively
accurately
route
from
isolation
also
context
bioengineering