Journal of Extracellular Vesicles,
Год журнала:
2024,
Номер
13(11)
Опубликована: Ноя. 1, 2024
Abstract
Extracellular
vesicles
(EVs)
have
gained
significant
attention
as
pathology
mediators
and
potential
diagnostic
tools
for
neurodegenerative
diseases.
However,
isolation
of
brain‐derived
EVs
(BDEVs)
from
tissue
remains
challenging,
often
involving
enzymatic
digestion
steps
that
may
compromise
the
integrity
EV
proteins
overall
functionality.
Here,
we
describe
collagenase
digestion,
commonly
used
BDEV
isolation,
produces
undesired
protein
cleavage
EV‐associated
in
brain
homogenates
cell‐derived
EVs.
In
order
to
avoid
this
effect,
studied
possibility
isolating
BDEVs
with
a
reduced
amount
or
without
any
protease.
Characterization
isolated
mouse
human
samples
(both
female
male)
revealed
their
characteristic
morphology
size
distribution
both
approaches.
show
even
minor
induces
‘artificial’
proteolytic
processing
key
markers,
such
Flotillin‐1,
CD81,
cellular
prion
(PrP
C
),
whereas
avoiding
treatment
completely
preserves
integrity.
We
found
no
major
differences
mRNA
content
between
non‐enzymatically
enzymatically
BDEVs,
suggesting
same
populations
are
purified
Intriguingly,
lack
Golgi
marker
GM130
signal,
referred
contamination
indicator
(or
negative
marker)
preparations,
seems
result
rather
than
its
actual
absence
samples.
Overall,
non‐enzymatic
is
possible
avoids
artificial
pruning
while
achieving
an
high
yield
purity.
This
protocol
will
help
understand
functions
associated
near‐physiological
setting,
thus
opening
new
research
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.
Journal of Extracellular Vesicles,
Год журнала:
2024,
Номер
13(6)
Опубликована: Июнь 1, 2024
Abstract
Isolation
of
neuron‐derived
extracellular
vesicles
(NDEVs)
with
L1
Cell
Adhesion
Molecule
(L1CAM)‐specific
antibodies
has
been
widely
used
to
identify
blood
biomarkers
CNS
disorders.
However,
full
methodological
validation
requires
demonstration
L1CAM
in
individual
NDEVs
and
lower
levels
or
absence
EVs
from
other
cells.
Here,
we
multiple
single‐EV
techniques
establish
the
neuronal
origin
determine
abundance
L1CAM‐positive
human
blood.
epitopes
ectodomain
are
shown
be
co‐expressed
on
single‐EVs
proteins
β‐III‐tubulin,
GAP43,
VAMP2,
which
increase
parallel
enrichment
EVs.
Levels
carrying
VAMP2
β‐III‐tubulin
range
30%
63%,
contrast
0.8%–3.9%
L1CAM‐negative
Plasma
fluid‐phase
does
not
bind
single‐EVs.
Our
findings
support
use
as
a
target
for
isolating
plasma
leveraging
their
cargo
reflecting
function.
Advanced Science,
Год журнала:
2024,
Номер
unknown
Опубликована: Ноя. 5, 2024
Abstract
Small
extracellular
vesicles
(sEVs)
act
as
a
critical
mediator
in
intercellular
communication.
Compared
to
sEVs
derived
from
vitro
sources,
tissue‐derived
can
reflect
the
vivo
signals
released
specific
tissues
more
accurately.
Currently,
studies
on
role
of
cochlea
have
relied
studying
sources.
This
study
evaluates
three
cochlear
tissue
digestion
and
sEV
(CDsEV)
isolation
methods,
first
proposes
that
optimal
approach
for
isolating
CDsEVs
using
collagenase
D
DNase
І
combined
with
sucrose
density
gradient
centrifugation.
Furthermore,
it
comprehensively
investigates
CDsEV
contents
cell
origins.
RNA
sequencing
proteomics
are
performed
analyze
miRNAs
proteins
CDsEVs.
The
crucial
maintaining
normal
auditory
function.
Among
them,
FGFR1
may
mediate
survival
hair
cells
via
sEVs.
Finally,
joint
analysis
single
single‐cell
data
is
utilized
trace
cellular
origins
results
show
different
types
secrete
amounts
CDsEVs,
Kölliker's
organ
supporting
most.
findings
expected
enhance
understanding
cochlea.
Journal of Extracellular Vesicles,
Год журнала:
2025,
Номер
14(1)
Опубликована: Янв. 1, 2025
ABSTRACT
Extracellular
vesicles
(EVs)
can
be
isolated
and
purified
from
cell
cultures
biofluids
using
different
methodologies.
Here,
we
explored
a
novel
EV
isolation
approach
by
combining
superabsorbent
polymers
(SAP)
in
dialysis
membrane
with
size
exclusion
chromatography
(SEC)
to
achieve
high
concentration
purity
of
EVs
without
the
use
ultracentrifugation
(UC).
Suspension
HEK293
cells
transfected
CD63
coupled
Thermo
Luciferase
were
used
quantify
yield
purity.
The
500
mL
conditioned
medium
volume
was
initially
reduced
pressure
ultrafiltration,
followed
UC,
SAP
or
centrifugal
filter
unit
(CFU).
Using
either
these
methods,
concentrated
final
approximately
1
mL,
retained
functionality.
yield,
quantified
luciferase
activity,
highest
UC
(70%–80%),
(60%–70%)
CFU
(50%–60%).
Further
purification
performed
iodixanol
density
cushion
(IDC)
SEC
(Sepharose
CL‐2B
6B,
10
20
columns).
Although
IDC
Sepharose
(10
mL)
achieved
yields,
slightly
higher
(30%)
IDC.
In
conclusion,
is
an
alternative
efficient
way
isolate
UC.
Journal of Extracellular Vesicles,
Год журнала:
2025,
Номер
14(1)
Опубликована: Янв. 1, 2025
ABSTRACT
Microglial
phagocytosis
of
haematomas
is
crucial
for
neural
functional
recovery
following
intracerebral
haemorrhage
(ICH),
a
process
regulated
by
various
factors
from
within
and
outside
the
central
nervous
system
(CNS).
Extracellular
vesicles
(EVs),
significant
mediators
intercellular
communication,
have
been
demonstrated
to
play
pivotal
role
in
pathogenesis
progression
CNS
diseases.
However,
regulatory
endogenous
EVs
on
phagocytic
capacity
microglia
post‐ICH
remains
elusive.
Utilising
multi‐omics
analysis
brain
tissue‐derived
proteomics
single‐cell
RNA
sequencing,
this
study
identified
that
bone
marrow‐derived
macrophages
(BMDMs)
potentially
enhance
microglial
via
ICH.
By
blocking
BMDMs
reducing
ARG1
BMDM‐derived
EVs,
we
facilitate
erythrophagocytosis
delivering
post‐ICH.
EVs‐carried
was
found
augment
promoting
RAC1‐dependent
cytoskeletal
remodelling
microglia.
Collectively,
research
uncovers
an
communication
pathway
mediated
This
provides
novel
paradigm
EV‐mediated
mechanisms
suggests
promising
therapeutic
potential
treatment
Journal of International Medical Research,
Год журнала:
2025,
Номер
53(2)
Опубликована: Фев. 1, 2025
Early
diagnosis
and
timely
management
are
critical
for
determining
disease
outcomes
prognoses.
To
date,
certain
methods
developing
disease-specific
biomarkers
have
been
reported;
however,
strategies
musculoskeletal
biomarker
development
rarely
studied.
Recent
studies
highlighted
the
potential
application
of
extracellular
vesicles
(EVs)
as
biomarkers.
EVs
encapsulate
proteins,
lipids,
messenger
RNAs,
microRNAs
derived
from
their
cellular
origin;
these
constituents
remain
stable
within
can
traverse
blood–brain
barrier.
Because
distinctive
characteristics,
actively
investigated
diagnostic
tools
various
conditions,
including
cancer,
inflammatory
diseases,
disorders.
Although
many
advantages
development,
they
not
yet
fully
researched
in
context
pathologies.
The
current
review
aimed
to
highlight
biomarkers,
summarize
processes
EV
discuss
limitations
future
perspectives
