From www.bloodjournal.org by guest on June 18, 2017. For personal use only.
Decreased
Membrane
Fluidity
Promyelocytic
During
Leukemia
By Stephen
The
human
(DMSO).
promyelocytic
phorbol
differentiation
5M.
nitrogen
cavitation.
ratio
ratio
process
to
were
bulk
this
cholesterol
in vitro.
membrane
cellular
contents
property
closely
core
regulated
processes
outside
studies
in
influence
coworkers
leukemia
undergoing
signals
in the
dimethylsulfoxide
proceeds
tumor
reflects
of
along
promoter,
(TPA)
induces
phages.7
Both
to influence
the
is
that
cells.’2
cell
a
forms
whether
DMSO
differentiation
influencing
the cell’s
Fluidity
in membrane
Among
the
by
growth
in
enviis a
in
to be
cellular
signals
from
Indeed,
recent
of membrane
and
vitro
environment.6
(DMSO),
Blood.
fluidity
Vol.
56,
No.
2 (August).
on day
cells
to
TPA,
lipid
leukemic
cells
cell
disruption
membrane
on
was
which
membrane
a small
No
these
and
to differentiate
cells
fluidity
increase
in
changes
in
became
occur
do not
1.6-
cytochemical
composition.
after
x
with
3 following
extracted
there
1 .6
probe
day
and
from
composition
process
(TPA).
morphological
5, and
promote
the
1 3-acetate
the
in phospholipid
in membrane
in
this
fluidity
response
that
might
Culture
human
Robert
Dr.
were
glass
during
themselves
the
appear
with
.tg/ml)
Laboratories,
determined
presence
of 2%
seeded
cal
Reagent)
was
added.
to
In contrast,
the
I 3-acetate
pended
2 mM
were
EDTA
From
determine
to
define
occur
on
cells
were
Pa.)
performed
salt
Island,
N.Y.).
Ill.)
University
ofO.5
to the
the
cell
Section,
Center,
were
resuspH
7.2
M sucrose,
cell
suspension.
cell
psi of nitrogen
to
were
Island
HEPES
in a pressure
800
Hematology-Oncology
cells
volume
dropwise
immediately
and
2 mM
cavitation
using
were
Cells
(Grand
The
and
Equal
added
Centrifuge)
solution
MgCI2,
cells
.5 x 106 cells/mI).
for
lysate
Department
(Parr
1 5 mm.
to
of
of Pennsylvania.
a final
Medi-
Philadel-
Pa.
in part
by Grants
AM-15441
and
CA-l#{243}52Ofrom
the
NIH.
the
by
changes
during
RC-3
balance
mM
was
added
cine and the Cancer
phia.
Warrington,
at 4#{176}C.
HL-60
(Sorvall
by nitrogen
Moline,
was
the
out
( I-I
0.5
7.5
lysed
Co.,
cells
Analyti-
Wright-Giemsa.
phase
Grand
pH
then
Instrument
with
of 2 x iO cells/mI.
HEPES
Cells
viabilin the
(Baker
suspensions
carried
g
M NaCI,
density
DMSO
were
Hanks’
Company,
at a cell
(100
and
to differentiation,
growth
I 20
with
in 0.14
Supported
to
at
twice
and
serum
Isolation
procedures
centrifuged
number
(Polysciences,
stained
at logarithmic
Biological
Na2
preparations
calf
in a hemacytometer
1 .25%
of cell
from
Md.)
streptomycin
Cell
induction
and
obtained
1 5% fetal
and
Va.).
For
counts
Membrane
were
U/mI),
M TPA
or 1.6 x l0
were
(Bethesda,
1640,
the cells
blue.
at 2 x iO cells/mI
All isolation
washed
by counting
slide
harvested
(100
McLean,
trypan
cells
Institute
in RPMI
penicillin
Differential
Plasma
Cancer
culture
supplemented
were
leukemia
of National
ity were
METHODS
Conditions
in continuous
(Flow
AND
promyelocytic
Gallo
kept
In the presence
differentiation
their
effect
leukemia
and
and
HL-60
on cytospin
differentia-
to differentiate
to macroTPA9
have been reported
of natural
or artificial
TPA
exert
promyelocytic
of differentiation.
process
prepared
sulfoxide
that
during
observed
with
change
HL-60
first
5. coincident
37%
significant
Cells
motion
It tends
well characterized.
study
was
undertaken
membrane
occur
of
was
liposomes
dimethyl
following
in
increased
but
have
described
a human
cell line that
is capable
of
HL-60
cells
DM508
and
the
fluidity
or
fluidity
MATERIALS
a barrier
molecular
myeloid
pathway.
tetradecanoyl-phorbol
of
agents
polarization
fluidity
on day
of
of
whether
sedimentation
fluorescence
stimulate
membranes
in vitro.
However,
the signals
for differentiation
and the way they are perceived
by cells are not
well understood.
Similarly,
the structural
changes
that
occur
in the surface
membrane
during
differentiation
have not been
The
present
density
highly
water
membranes.
differentiation
chemical
no
presence
or tetradecanolyl-phorbol
observed
changes
agents
extracellular
thought
to be controlled
of the cell is differentiation.
have indicated
that alternations
fluidity
may
tion
Gallo
and
promyelocytic
with
the
determine
in membrane
membrane
was
significant
signals
from
of the cell.
that
hydrophobic
were
1 8 hr of exposure
that
However.
and
senses
and transmits
ronment
to the interior
the
the
a maximum
acids,
changes
1 .25%.
in
in
to
fluidity.
also
structural
by
phospholipid
A. Cooper
differentiation
sucrose
decrease
fluidity
fatty
during
by
of Human
in Culture
undertaken
whether
cells
reached
to
demonstrate
were
assessed
membrane
HE
SURFACE
MEMBRANE
specialized
structure.
It not only
between
of
was
unsaturated
undergoes
of DMSO,
from
Cells
p and Richard
to assess
progressive
and
in
observed
studies
of differentiation
to perturb
T
to DMSO.
of membrane
fluidity
These
A
changes
of saturated
membrane
adherent.
cells
HL-60.
presence
isolated
fluidity
H.C.
Studies
and
in the
(DPH).
Similar
The
grown
Membrane
of HL-60
line.
agents.
fluidity
were
.3,5-hexatriene
exposure
the
were
membranes
differentiation.
cell
other
membrane
Cells
Surface
diphenyl-1
lipids.
and
influence
of differentiation.
10
leukemia
esters,
Differentiation
Submitted
Oncology
Spruce
the
November
Address
©
reprint
Section.
Street,
30, /979;
requests
Hospital
to
ofthe
Philadelphia.
1980 by Grune
accepted
Richard
& Stratton.
Pa.
University
April
A.
8, 1980.
Cooper.
Hematology-
ofPennsylvania,
3400
19104.
inc.
0006-497l/80/5602-00l2$Ol.00/0
1980
227
From www.bloodjournal.org by guest on June 18, 2017. For personal use only.
228
IP AND
concentration
I 5 mm.
of 0.5
The
buffer
and
solutions
membranes
were
HEPES,
pH
37%
sucrose
solution
was
centrifuged
lysates
were
resuspended
before.
at
100,000
The
resuspended
7.2.
The
a cellulose
at
lysis
supernatant
sucrose
solution
The
pelleted
solution
with
was
overlayed
nitrate
centrifuge
100,000
g in a Beckman
2
on a
tube.
The
was
serum
of plasma
measured
by
cytochrome-C
to the
determined
albumin
method
and
of
reductase
method
by the
Lipid
Extraction
was
g.
procedure
homogenates
Kidwei
and 5’-nucleotidase
et
prior
Extraction
of
membrane
using
from
suspension
the
method
extracted
lipids
were
and
assayed
depolarization
not
Dispersions
Dyer.’4
for
of membrane
the
nitrogen
(Long
Island,
Fatty
Acid
Analysis
All organic
solvents
tilled
calculated
from
to the
Fatty
by
Hewlett
to
ionization
described
by
Chrom
State
aqueous
chloroform:methanol
and
the
plasma
chloroform:methanoltotal
College,
rising
30
esters
lipid
of the
al.’7
for
a Heat
and
acid
using
Phospholipid
used
for
a
held
Whatman
(Whatman
lipids
mixture
applied
before
After
being
with
cupric
was
then
analyzed
(model
ment
after
plasma
by thin-layer
plate.
developing,
The
sprayed
dried
the
with
for
20
using
a
equipped
Westwood,
mm
plate
with
emitted
‘I
‘I
according
light
(P)
parallel
to the
and
formula.
i
+
dried
spray
phosphoric
10 C
Inc.,
acids.
The
of sample,
of
HL-60
cells
I). The activity
growing
in
continuous
of NADH-cytochrome-
membrane
fraction.
Membrane
Fluidity
Changes
During
reticuin the
succinic
purified
Cellular
Differentiation
Nitrogen
peaks
The
was made
maturation
of HL-60
cells
is maximal
5-7
days
the addition
of 1 .25% DMSO.6
Membrane
fluidity was measured
in HL-60
cells during
this period
of
differentiation
using
the
hydrophobic
fluorescent
probe,
DPH
(Fig.
I). No observable
differences
were
after
membrane
by chloon
preabsorbent
plates
5 gg of phosphophase
for
5 mm
reagent
acid.’9
I 30#{176}C.
Phospholipid
according
from
plasma
membrane
fraction.
No mitochondrial
dehydrogenase
activity
was detectable
in the
Lab.
10 mm.
or
dual
as
Silar
fatty
mobile
Schoeffel
dual-beam
with a density
computer
Ni.)
of
by
polarization
C reductase,
an enzyme
present
in endoplasmic
lum, was increased
by an average
of I .4-fold
Pa.)
was
in the
was
The
in 8%
at
intensity
of excitation
as monitored
was
chromatography
developing
reagents.
30 g/liter,
Fluorescence
markers
were used to assess
the purity
membrane
preparation.
5’-Nucleotidase
(Table
culture
computerized
injection
at 190#{176}Cfor
from
separated
to the
acetate,
by
3000)
Corp.,
were
10%
of chloroform:methanol:acetic-acid:water
25:15:4:2.
lipids
Science
of
mm
2#{174}-ithick 10 x 10 cm LHP-K
Inc., Clifton,
Ni.).
Approximately
was
redis-
Avondale,
(Applied
quantitation
extracted
were
at 37#{176}C
to optical density of less than 0.1
polarization.
Light scattering
the activity
in the whole
cell homogenate,
a 15-fold
enhancement
of (Nat
K)ATPase
activity
in 6 separate
plasma
membrane
preparations
isolated
from either
DMSO-induced
HL-
were
analysis
A 6 ft x 2 mm,
column
at I 55#{176}C
for 0.5
mixtures
at a protein
incubated
with
was
there
Analysis
roform:methanol
dispersion
Purification
Enzyme
plasma
pared
under
Systems
equipped
flow was 25 mI/mm.
Identification
of the various
by comparison
with known standards.
Phospholipid
the
5 mm
extracted
Fatty
Packard,
mesh
to 190#{176}C
at 1#{176}C/mm, and
was
mixing
DPH
preparation
mixture
diluted
activity,
a commonly
used plasma
membrane
enzyme
marker,
was
not detectable
either
in isolated
cell
membranes
or whole
cell
homogenates.
However,
(Nat
K)-ATPase
activity
was present.
As com-
phos-
by sonicating
W
grade
chromatograph
Blough.’8
was
held
analytical
(Hewett
and
Pa.)
was
at
chromatography
detectors
Baron
prepared
at 4#{176}C
using
et
gas
Q 100/120
temperature
were
methyl
5830A
the
plane
and
RESULTS
sonifier.
Litman
gas-liquid
membrane
the
of fluorescence,
P=
60 cells
acid
Packard
flame
W375
used
of the diluted
of samples.
was
The
extracts
saline
model
according
performed
Gas
N.Y.)
in glass.
prepared
were
lipid
in phosphate-buffered
dilutions
M
vigorous
accord-
with
cholesterol’3
lipids
chloroform:methanol
cause
of serial
NADH-
16
dried
and
itself
perpendicular
dehydrogenase
of
performed
Bligh
tzg/ml
I0
with
et al.’3
aliquots
was
of
volume
were diluted
of fluorescence
Analysis
lipids
One
for 30 mm. Samples
for measurements
did
at
saline
of plasma
of 250
Membrane
and
tetrahydrofuran
to use.
to I volume
concentration
was
et
in
phosphate-buffered
were
al”
were measured
Succinate
of DeDuve
of Lowry
dissolved
with
was added
at
K)-ATPase
+
cell
and
et al.’2
method
(Na
whole
Abdul
of Avruch
assayed
by the
as standard.’0
membranes
the
Wisc.)
means
concentration
bovine
activities
ing
at
Measurements
Measurement
of fluorescence
polarization
were performed
with
an Elscint
MV- I microviscosimeter
(Elscint
Corp.,
Hackensack,
N.J.) as reported
previously.2122
The fluorescent
probe, I ,6-diphenyl- 1,3,5-hexatriene
(DPH)
(Aldrich
Chemical
Co., Milwaukee,
immediately
Essays
Protein
al. using
Polarization
1000-fold
S.W.
50.1 rotor (Beckman
L-65
centrifuge).
The membrane
fraction
the interface
was removed
and resuspended
with I mM HEPES
pH 7.2. The sample
was then centrifuged
for 90 mm at 100,000
Pellets were resuspended
in appropriate
solutions.
Biochemical
Fluorescence
g for
of the
combined
in 25%
for 90 mm
at 5500
volume
g for 60 mm.
homogenized
in
centrifuged
in the same
as
centrifuged
mM
sample
Cell
were
recentrifuged
were
crude
mM.
pellets
COOPER
ratio
was
made
The
plate
spots
et
of
at 60#{176}C
were
spectrodensitometer
(Schoeffel
Instru-
to Touchstone
a
seen
during
the
DMSO.
Thereafter,
DPH
increased,
first
2 days
after
the fluorescence
indicative
of a decrease
the addition
polarization
of
of
in membrane
fluidity.
The
maximum
change
in fluidity
was
at 5 days,
when
over 90% of the cells were
differentiated
as judged
by the presence
of myelocytes,
metamyelocytes,
and bands
on Wright-stained
smears.
observed
Thus,
plasma
the morphological
The
membrane
membrane
fluidity
changes
appearance
of differentiated
fluidity
of
HL-60
cells
paralleled
cells.
was
also
From www.bloodjournal.org by guest on June 18, 2017. For personal use only.
MEMBRANE
FLUIDITY
IN
Table
HL-60
CELLS
1 . Enzymatic
229
Activity
of Whole
HL-60
Cell Homogenates
and of Membrane
Fractions
of HL-60
Membrane
Whole
(Na-Ki-ATPase
OD
3
at 700
nM/mg
Cytochrome-C
reductase
2
2
(24. 1 -37.0)
5.2
(5.5-5.7)
0.28
protein/hr
1)
5.6
3.8
Cells
31.4
(25.2-40.
(3.5-4.2)
dehydrogenase
zmoIe/mg
DMSO-lnduced
32.1
(1 .82-2.40)
.tmoIe/mgprotein/hr
Succinate
Growing Cells
2.14
protein/hr
Fraction
Cell
Homogenate
n
Cells
(4.5-6.9)
0
0
(0.22-0.35)
‘Range.
analyzed
after
exposure
of the
TPA,
adhere
a phorbol
ester.
Because
to the plastic
substrate
hr
exposure,
of
studies
of
cells
to
1.6
x
108
TPA
induces
cells
after approximately
membrane
fluidity
carried
out for only the first I 8 hr. Membrane
at 18 hr was identical
in HL-60
cells, whether
M
lipids,
to
18
measured
were
fluidity
or not
they
the
been
As
fluorescence
polarization
in liposomes
prepared
extracted
from the isolated
with
seen
of l-IL-60
were less
whole
of
DPH
from lipids
membrane
membranes,
the
cells 5 days
after
fluid than
the lipids
was
that had
fractions.
membrane
induction
extracted
with
from
lipids
DMSO
HL-60
had been exposed
to TPA (Fig.
I).
Figure
2 shows
the fluorescence
polarization
of
DPH in the plasma
membrane
fraction
of HL-60
cells
5 days after exposure
to DMSO.
Membranes
isolated
from
differentiated
cells
were
less
fluid
than
the
cells in continuous
culture
(Fig. 3).
In various
biologic
systems,
membrane
fluidity
has
been found to be influenced
by the relative
amounts
of
cholesterol
and phospholipid,22
by the specific
classes
of phospholipids,23
and by the degree
of saturation
of
membranes
indicated
the fatty acids
characteristics
present
in phospholipids.24
of lipids
were analyzed
from
from HL-60
cells in continuous
by the significantly
higher
values
cence
polarization
tures.
As
over
the
elutes
made
cells
at all
tempera-
in membranes
was
observed
and
transition
of
in the
fluorescence
presence
the
polarization
of 1 .25%
DMSO.
No
effect was observed
when this concentration
of DMSO
was added
to isolated
membranes.
Similarly,
5.0%
DMSO
had
membranes.
Analysis
To
no effect
on the
of Membrane
determine
membrane
of red
cell
ghost
were
the
due
observed
to changes
changes
HL-60
was
relative
HL-60
cells
cells
5
of
in continuous
days
no statistically
quantities
These
three
quantitatively
after
significant
the
various
culture
exposure
to
difference
in
phospholipids
present
before
and following
differentiation
(Table
2).
An analysis
of the fatty
acyl chains
present
in the
membrane
lipid
extract
is shown
in Table
3.
Compared
with
HL-60
cells
in continuous
culture,
DMSO-differentiated
HL-60
cells had slightly
more
saturated
fatty acids with a small decrease
in 1 6: 1 and
Lipids
whether
fluidity
fluidity
from
DMSO.
There
range
of lO#{176}-45#{176}.
Since
DMSO
during
the process
of membrane
measurements
also
induced
no phase
temperature
from
cells
isolation,
were
in the
expected,
culture,
as
for fluores-
in
in membrane
18:1. The cholesterol/phospholipid
of the plasma
membrane
fraction
in
growing
continuously
(C/P)
increased
HL-60
cells
to
mole
from
ratio
0.35
0.48
after
U
035
027
0.
0
.2
0
030
0.26
0.25
DMSO
Induced
HL-60
0
0.
. a
0
I
025
2
Time
3
4
-60
HL
5
(days)
to
20
30
Temperature
Fig. 1 .
Time course of changes
in membrane
fluidity following
exposure
to DMSO
(#{149})
or TPA (A). The earliest
changes
in fluidity
were observed
at 60 hr and fluidity
was maximally
affected
at 5
days. Values
at zero time were
identical
to values
obtained
in
control
cultures
without
DMSO
or TPA.
40
50
1’ C)
Fig. 2.
Fluidity
of DPH in membranes
isolated
from continuously
growing
HL-60
cells and from
HL-60
cells 5 days after
induction
with DMSO.
The membrane
fluidity
of induced
cells was
decreased.
as indicated
by higher values for P at all temperatures.
From www.bloodjournal.org by guest on June 18, 2017. For personal use only.
230
IP AND
Table
035
3.
Fatty
A cyl Chains
of Plasma
Percent
Membrane
of Total
Fatty
COOPER
Lipids
Acids
0.
DMSO-Induced
0
Acyl
030
HL-60
Group
14:0
DMSO Induced
HL-60
16:0
t_ .60
020
0
20
--
40
30
Teryerature
(SC)
Fig. 3.
Fluidity
of DPH in liposomes
prepared
from the lipid
extract
of membranes
isolated
from continuously
growing
HL-60
cells and from HL-60 cells 5 days after induction
with DMSO.
Lipid
fluidity
was decreased
after DMSO induction.
6.1
26.8
16:1
025
HL-60
3.9
26.0
5.1
3.7
18:0
22.4
26.0
18:1
15.4
10.6
18:2
2.3
1.8
20:1
20:2
20:3
0.7
0.4
+
22:0
0.7
0.5
+
20:4
5.9
7.0
20:5
1.8
1.9
22:1
2.4
2.9
22:5
2.5
2.7
22:6
24:1
3.2
4.2
24:0
1.5
0.7
+
Unsaturated/
differentiation.
ciated
0.020.
This
37%
with a change
When
expressed
of 0.59
poise
in C/P
was
from our
coworkers,2’
(from
of
as
laboratory22’23’25
this
represents
2.73
poise
before
saturated
asso-
in fluorescence
polarization
in terms
of microviscosity,
in previous
publications
from
Shinitsky
and
change
increase
and
a
differen-
tiation
to 3.33 poise at day 5). In our previous
studies
with red cells, a 37% increase
in membrane
cholesterol
above
baseline
values
resulted
in an increase
in micro-
viscosity
of 0.60 poise,22 and a 37% increase
in
cholesterol
led to an increase
in microviscosity
poise.25
Thus,
the change
in C/P
in HL-60
differentiation
was of a magnitude
that could
for the observed
changes
in fluidity
observed
DPH.
platelet
of 0.52
cells on
account
with
These
is
studies
demonstrate
associated
with
membrane
increase
Table
a
fluidity
and that
this
in the cholesterol/phospholipid
2.
Plasma
Induced
Membrane
result
mole
Composition
Growing
Leukemia
differentiadecrease
is the
Phospholipid
and Continuously
HL-60
in
of an
ratio
of DMSO-
ethanolamine
serine
phosphatidyl
Phosphatidyl
±
1.4
33.7
±
Inositol
Sphingomyelin
19.1
±
0.5
18.7
±
33.0
±
1.1
35.7
±
0.2
0.6
10.3
±
0.5
12.0
±
2.3
Cholesterol/phospholipid
0.35
(mole/mole)
Results
are
an
standard
deviation.
individual
experiment.
average
Triplicate
of
three
TLC
DMSO-induced
separate
measurements
cells
undergo
were
0.48
pairs
of
were
studied
a
decrease
fibroblasts
decreases
fluidity
of chicken
embryo
an
increased
C/P
when
these
cells
become
in vitro.29
It is likely
that
changes
in
lipid
composition
and membrane
fluidity
the
enzymatic3#{176} and
membrane
modify
Similarly,
membrane
differentiation.28
fluidity
due to
confluent
in
HL-60
also
during
the mem-
cells as they assume
their
Do inducers
of cellular
through
primary
transport
properties3t’32
new physiologic
differentiation
changes
role.
evoke
in membrane
of phospholipid
experiments
made
for
on day
5.
membrane
of
their
fluidi-
concentratempera-
vesicles,
suggesting
that
DMSO
fluidity.8
We
have
confirmed
these studies
in our laboratory
but have noted that,
at
the concentrations
of DMSO
utilized
to induce
differentiation,
these changes
in vesicles
are not measurable
2.0
+
choline
DMSO-induced
on day 5.
immaturity
rather
than of malignancy.27
Like
cells,
Friend
erythroleukemia
cell membranes
decreases
HL-60
37.6
experiments.
than
that of mature
polymorphonuclear
leukocytes.26
Recently,
Klock
and
Pieprzyk
demonstrated
that
normal
immature
bone marrow
cells also have a low
C/P.
suggesting
that a low C/P
is a manifestation
of
ture
DMSO-lnduced
Phosphatidyl
37.5
separate
ty? Previous
studies
have shown
that high
tions of DMSO
increase
the phase transition
To tel Phospholipids
HL-60
studied
of two
and in the degree
of fatty acyl chain
saturation
within
the cell membrane.
These
observations
are consistent
with data reported
by Gottfried
showing
that the C/P
mole ratio
of myeloblasts
in acute
leukemia
is lower
signal
Promyelocytic
Cells
Percent
Phosphatidyl
cells were
brane
that myeloid
progressive
an average
are
DMSO-induced
DISCUSSION
tion
Results
41.9
±
each
(‘p and Cooper,
unpublished).
Moreover,
no change
in
HL-60
membrane
fluidity
was observed
using
these
concentrations
of DMSO.
Phorbol
esters
have
also
been reported
to effect membrane
fluidity.
For example, TPA
has been
noted
to increase
the membrane
fluidity
of
rat
embryo
cells9
and
of
a
variety
of
From www.bloodjournal.org by guest on June 18, 2017. For personal use only.
MEMBRANE
FLUIDITY
lymphoid
IN
cells33
above
the
at
range
noted
only
and,
among
those
In
subpopulation.
ity
that were
of the
leading
231
CELLS
that
concentrations
utilized
the case
in those
tiation.
HL-6O
to induce
are
HL-60
cell
lines,
This suggests
observed
may
only
in
the
to detect
fluidity
any
HL-60
structure
of
membrane
adherent
change
in the
membrane
terize
the
be
emphasized
that
process
measurements
probes
such
bulk
fluidity
the
as
DPH
of
report
the
entire
of
fluidity
in HL-60
cells at
differentiation.
Rather,
the
observed
appear
to charac-
ofcellular
probe
of
previous
differentiation.
ACKNOWLEDGMENT
The
studies.
It should
for
alized effects on membrane
concentrations
that induce
changes
in fluidity
we have
an early
signal
study
we were
utilizing
the same
was utilized
in these
with
values
membrane,
and local changes
in fluidity
that might
be
of great
functional
importance
may not be detected.
Thus,
although
such local changes
may exist,
it does
not appear
that either
DMSO
or TPA induces
gener-
that the changes
in fluidhave been a manifestation
early
cells
that
fluidity
average
lymphoid
cells,
changes
were
cell lines that
became
adherent
of
TPA
effect
on cells rather
than
to that effect.
In the present
unable
membrane
at or just
cell differen-
authors
Penny
gratefully
acknowledge
the
technical
assistance
of
Shames.
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1980 56: 227-232
Decreased membrane fluidity during differentiation of human
promyelocytic leukemia cells in culture
SH Ip and RA Cooper
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