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A Method
for Simultaneous
Study
Morphology,
and Immunologic
Mitotic
Cells
in Hematologic
S. Knuutila,
By L. Teerenhovi,
Ekblom,
M.
L. Rossi,
of the Karyotype,
Phenotype
of
Malignancies
Borgstr#{246}m, J.K. Tallman,
G.H.
L. Andersson,
and A. de Ia Chapelle
problem
in the cytogenetic
analysis
of hematologic
neoplasms
has been an inability
to identify
the cell from
which
the chromosomes
were
obtained.
We describe
a
procedure
that allows simultaneous
analysis
of karyotype
and cell cytology
in mitotic
cells. The method
differs from
conventional
cytogenetic
analysis
in that after mild hypetonic treatment.
the cells are cytocentrifuged
onto glass
slides.
In mitotic
cells. this procedure
often
results
in
adequate
spread of the chromosomes
within the intact cell
membrane.
The cytoplasmic
structure
also remains
intact.
so that cytologic
preparations
are of good quality. Morphologic and immunologic
identification
of mitotic
cells can be
done using routine
hematologic
stains. such as Giemsa
or
Sudan black B. and various antisera
using immunofluorescence
techniques.
The chromosomes
can
be simultaneously analyzed
either without
banding on slides stained
with
Giemsa
or with Q-banding
on slides stained
with
immunofluorescence
techniques.
Identification
of numerical and structural
karyotype
aberrations
thus is possible in
morphologically
identified
cells.
C
variety
A major
ELL
LINEAGES
cytogenetic
quently,
cannot
methods
in bone
marrow
types
undergo
lineages
are
mitosis,
represented
chromosomes
are
several
found,
respects.
it cannot
be identified
currently
(BM),
one
studied.
This
the
Conse-
cells
of several
where
does
among
with
available.
not know
what
cell
the mitoses
whose
is a major
drawback
more than one cytogenetic
abnormality
is found,
not possible
to know
which
abnormality
belongs
which
cell type. Third,
if a mixture
of cytogenetically
netically
normal
cells
are
some
of the leukemic
abnormal
and others
karyotypes
are found,
the
leukemic
whether
the
normal
nonleukemic.
lineages.
These
situacytoge-
can
when
In this
Cells
aspirated
to vials
we show
that
identification
several
can
be
gradient
Falcon
humidified
following
three
hours.
medium,
0.8
for
with
a
into
50
added
in a final
occasional
I 2 cytocentrifuge
were
chambers
and
5%
in
NY)
to
in 2 mL
of
solution
KCI,
10 mmol/L
and
10 mmol/L
standing
the suspension
(Cytospin,
was
two
a hypotonic
After
for five
was
Shandon
a
The
CO2.
Island,
CaCI2,
of 4 mL.’
mixing,
corn, England) and centrifuged
air-dried
overnight,
the slides
above in sterile
suspended
of
1 mmol/L
gentle
1640.
overnight
5 mmol/L
volume
Fine
RPM!
for an additional
2 mL
glycerol,
with
Grand
I 0 cells
MgCl2,
7.0)
with
was
mmol/L
mmol/L
(pH
minutes
slides,
by one-step
(Pharmacia
air
incubated
trans-
penicillin
(0.29 mg/mL),
incubated
(GIBCO,
were
isolated
twice
95%
or
with
described
were
#{176}C
with
the flasks
to which
of
washed
flasks
37
To prepare
consisting
NaC1,
and
were
in the medium
Colcemid
cg/mL)
(PB)
in Ficoll-Hypaque
The
at
morning,
(0.4
the
flasks.
blood
supplemented
cells
and
show
Study
Cytochemical,
L-glutamine
Nucleated
Sweden)
incubator
added
1640
centrifugation
We
and
METHODS
g/mL),
(50
serum.
Uppsala,
25-mL
be
techniques
calf
cell.
method
or peripheral
The cells were resuspended
only
of the
RPMI
streptomycin
Chemicals,
a way has
the cells that
combined
BM
immuno-
mitotic
Cytogenetic
Morphologic,
containing
10% fetal
sucrose
article
from
and
same
AND
ofSlidesfor
With
Analysis
(50 U/mL),
or
the
MATERIALS
Preparation
Simultaneously
Immunologic
and
12
chromosome
of
the basic
principles
of its application.
density
normal
nonleukemic
questions
resolved
characterize
describe
examples
ferred
cells might
be cytogenetically
normal.
Fourth,
if only normal
the major
question
is whether
answered
and controversies
been found
to identify
and
are
it is
to
Alternatively,
cells
are
cytogenetically
cells karyotyped
represent
cell
is
if
for the morphologic
characterization
in
First,
if a cytogenetic
abnormality
be assigned
to a cell lineage.
Second,
normal
and abnormal
cells is found
(a common
tion in leukemias),
one cannot
tell whether
the
of techniques
logic
divided
Elliot,
Run-
at 400 rpm for ten minutes.
When
were ready to be used for further
analysis.
From
ment
the
Department
of Medicine,
of Medical
and
Genetics,
the
Transplantation
supported
by grants
the
Third
Laboratory,
DepartUniversity
Cytochemical
of Helsinki.
This
study
Foundation,
dation,
was
the Finnish
the Nordisk
Folkh#{228}lsan Institute
Submitted
Address
26. /983;
requests
Department
Helsinki,
I 984 by Grune
the Academy
Finnish
of Finland,
For
Cultural
Juselius
and
of
to
Dr
Medical
the
June
L.
Genetics,
were
4. 1984.
Teerenhovi.
University
Haartmaninkatu
of
Giemsa
3,
water.
Inc.
cytologic
would
Giemsa
cytic
Finland.
& Stratton,
routine
alternative
Foun-
staining
methanol
accepted
0006-4971/84/6405-0027$03.00/0
1116
the
the Sigrid
of Genetics.
Sept
Helsinki,
from
Society,
lnsulinfond,
reprint
SF-00290
©
Cancer
Staining
were
(49.5
washed
diluted
Sudan
cells.3
hematoxylin
be
mL)
in pure
with
black
Methods
analysis,
Giemsa
staining
May-Gruenwald-Giemsa
fixed
and
for
five
37%
methanol
and
water
B staining
was
a/Naphthyl
counterstaining
to ten
minutes
formaldehyde
buffered
acetate
(30
used
for
6.8),
used;
Slides
mL).
The
of
slides
30 minutes
in 5%
then
in tap
to identify
esterase
(ANAE)
minutes)
was
rinsed
early
granulo-
followed
used
to
identify
monocytes.4
Blood. Vol 64, No 5 (November),
an
for
in a mixture
(0.5
stained
(pH
was
stain.
1984: pp 1116-1122
by
From www.bloodjournal.org by guest on June 16, 2017. For personal use only.
SIMULTANEOUS
STUDY
Immunologic
The
OF KARYOTYPE
AND
MORPHOLOGY
1117
Methods
slides
were
fixed
for five minutes
but sometimes
The ANAE
in the methanol-formalde-
hyde solution
as above, or at - 20 #{176}C
in a solution
containing
50 mL
and four drops of formaldehyde.
After two washings with
1640 and 2% bovine serum albumin,
antibody
was applied
to the wet cells on the slides.
acetone
RPMI
directly
To
identify
prepared
from
erythroid
the
IgG
at a
used
with
RPMI
of 1:20.
F(ab)2
fragments
The
preparation
bovine
(TRITC)
was incubated
for
I 5 minutes
serum
with
and
and
once
of
CALL
the
lymphoid
I antiserum
containing
and
(eg,
case
Los Angeles).
lysozyme
were
2) were
identified
Pharmacia
Fine
with
antisera
Chemicals).
conjugated
with
wet
with
RPMI
RPMI
bovine
1640-2%
albumin
The
agent
some
was
antiserum
and
once
with
Chromosome
the
(Fig
IA,
Giemsa
above
B).
Sudan
When
banding
Sudan
black
stain
stained
black
used for routine
the
B stain
gave
was not required,
B were
examined
cytologic
chromosomes
for
both
analy-
without
a similar
slides
stained
cytologic
banding.
Preparations
in an alcohol
series:
ethanol.
chromosomes
The
described
stained
for Q-banding.
be studied
The
graphed
ASA)
minutes
The
simply
result
(Fig
with
Giemsa
IC).
state
and
or
karyo-
karyotype
filters
tively,
the TRITC-positive
be
exposure
antisera
could
and
TRITC
fluorescence
on
N2
for TRITC
mitotic
photographed
(Fig
in absolute,
restained
the filters
mitoses
could
each
were
70%,
for
destained
and
30%
Q-banding
immediately
ofTRITC-positive
by changing
a Leitz
with
Giemsa
as
et al.6
fluorescent
Q-banded
with
film
with
with
then
were
by Caspersson
Slides
scope.
five
stained
on
the
cells
same
cells
could
fluorescence
micro-
cells
photo-
positive
Kodachrome
and
be used
mitotic
in the
were
I 35-20
E2 for Q-bands.
and Q-banded
small
cells
lymphoblasts,
precluded
(400
Alterna-
chromosomes
photomicrograph
by
double
I D).
The
methods
ofCell
gave
Type
cytologic
preparations
of good
when
to
large
cell (Fig iF).
In cells of granulocytic
often
possible
from
the
with
and cell size and was
a nearby
nonmitotic
origin,
typical
identification
granulation
was
(Fig
1A),
cells
tural
and
detection
lymphocytes.
in 7/12
aberrations,
of
of mod-
promyelocytes,
cases
allocation
allowed
groups
the
We
of
have
adult
and
1/6
Giemsa
chromosomes
of numerical
and
eg, the Ph’ chromosome
to
certain
struc-
(Fig
1A, B).
To avoid excessive
overlapping
of the chromosomes
the
cultures
were treated
with Colcemid
for as long as two
to three hours.
This led to chromosomes
of medium
to
small
could
banding
size,
not
in which
small
always
be
after destaining
quality
deletions
or translocations
identified
with
certainty.
of Giemsa
or in conjunction
TRITC-conjugated
antisera
(Fig
structural
abnormalities
different
staining
methods
ing
methods
requiring
extensively
ase reaction.
counterstained
characteristics
by comparison
from
was
were frebest chro-
staining
on cytoplasmic
often helped
nonmitotic
obtained
myeloblasts,
large
mitoses
or
of chromosome
granulation
of CML
in blastic
crisis,
ALL.
was good,
conventional
very weak
no banding.
staining,
lymphocytes
of chromosomes
ANLL,
4/6 cases
cases of childhood
When
spreading
proved
Giemsa
chromo-
greatly
on
the mitotic
the chromosomes
Consequently,
the
eg,
erythroblasts,
and
obtained
analyzable
produced
usually
1 B, F). After
depended
within
cytoplasmic
were
size,
Giemsa
(Fig
the
preparations
erate
cells could easily be identified
as to cell lineage
(Fig 1,
B, E, F). In mitotic
cells, identification
had to be based
quality
for
the determination
(as in myelocytes),
fuzzy
(Fig IA).
moderate
RESULTS
Identification
antinot
immunofluorescent
Q-banding
such
as small
overlapping
even
Also,
with
Cytologic
identification
Identification
cell.
In
LI-type
mosome
banding
type.
With
of TRITC
The quality
of the preparations
degree
of chromosome
spread
heavy
quently
of Chromosomes
sis as described
For
of
Copenha-
often
banding.
use
allowed
simultaneous
identification
(Fig I D).
number.
Without
1D).
as the
1640.
Staining
(Fig
shown).
and
preparation
anti-rabbit
serum
anti-GPA
several
for direct
identification
series
were successfully
with
(Dakopatts)
was then applied to the cells, and the preparations
were
incubated
for I 5 minutes in a moist chamber. Slides were washed
once
of fluorescence,
)
with
in a moist chamber and
serum albumin.
Tetra-
swine
but chromosome
staining
only
the identification
of
aberrations.
as a source
with
as well.
mitotic
and B lymphocytes,
were used (examples
antikap-
(Dakopatts,
The
incubated for I 5 minutes with the antiserum
washed twice with RPMI
1640-2% bovine
methylrhodamine
polyvalent
gross numerical
With
TRITC
to be used
to identify
of monocytes,
antilysozyme,
kappa
or antilambda
antisera
Cells of B cell origin
identified
, and antilysozyme
pa,- antilambdagen,
series
(UCLA,
of the monocyte
lineage,
often
weak,
allowing
identified
washed
albumin
black
B had
was also used
antisera
(Table
I were used
cells.
Cells
of the erythroid
A (anti-GPA)
tetramethylrhodamine
chamber
1640-2%
cells,
of anti-glycophorin
with
in a moist
RPMI
of the
1640.
Cells
cells
fraction
dilution
anti-GPA-TRITC
once
origin
and conjugated
antiserum
were
the
cells
was
Sudan
reaction
difficult
ID),
gave
which
did
to be found.
that proved
counterstains
because
Q-
banding
permit
of
such
Table
I lists the
valuable.
Stainother
than
such
counterstains
staining
of the chromosomes
The only
method
of this
and
type
studied
was the a-naphthyl
acetate
esterWith this staining
method,
chromosomes
with
hematoxylin
were
occasionally
analyzable,
while
toluidine
stain the chromosomes.
blue
regularly
failed
to
From www.bloodjournal.org by guest on June 16, 2017. For personal use only.
1 1 18
TEERENHOVI
ET AL
Fig 1 .
Peripheral
blood preparation
for simultaneous
cytogenetic
and morphologic
study of case 1 . (A) A mitotic
cell with myeloid
granulation
(Giemsa
staining).
(B) A mitotic
cell without
myeloid
granulation
(Giemsa
staining).
(C) A Sudan-negative
mitotic
cell. A Sudan
positive
cell is indicated
by the arrow
(Sudan
black B staining).
(D) An anti-glycophorin-A-positive
mitotic
cell. Combined
staining
with
anti-glycophorin-A
coupled
with TRITC and quinacrine
in double exposure.
The bright red color of TRITC was deliberately
underexposed
at photography
to enhance
the visibility
of the quinacrine
banding.
A TRITC-negative
cell is indicated
by the arrow.
(E) Bone marrow
preparation
for simultaneous
cytogenetic
and morphologic
study of case 2. Two cells in mitosis.
The mitotic
cell on the left has slight
myeloid
granulation.
In both cells the karyotype
is 47.XX.
+ G.Ph’.
(F) Bone marrow
preparation
for simultaneous
cytogenetic
and
morphologic
study
from
a patient
with
acute
monocytic
leukemia
and karyotype
46.XY.
A mitotic
cell with
cytoplasm
resembling
neighboring
nonmitotic
cells. some of which (arrows)
have a clearly monocytoid
appearance
(Giemsa
staining).
From www.bloodjournal.org by guest on June 16, 2017. For personal use only.
SIMULTANEOUS
STUDY
OF KARYOTYPE
AND
MORPHOLOGY
1119
Applications
cytes.
Case
1.
A 68-year-old
woman
had
as having
chronic
myeloid
leukemia
phase.
At that time the karyotype
the standard
constitutional
ulated
years,
the
total
a differential
myelocytes,
mented
blood
was
phocytes.
The
cells
count
2%
blasts
any
was
+22q-,t(9;22)
bone marrow
tional
trypsin
arations
cells, was
diagnosed.
leukocyte
without
contained
these
normal.
In the
was
78.0
similar.
x 109/L
and
moderately
granulation
80% blast
with
10%
basophilic
(Fig
cells,
The
After
four
peripheral
5% promyelocytes,
30% stab and seg-
eosinophils;
had
the
lymcyto-
1 B). In the
morphology
karyotype
BM,
of
48,XX,+
19,
was seen in both peripheral
blood and
in all 22 mitoses
studied
with the convenGiemsa
banding
method
(Fig 2A). Prep-
for simultaneous
analysis
stained
(CML)
in chronic
was 46,XX,Ph’
with
count
of 53% blasts;
and metamyelocytes;
granulocytes;
plasm
which
diagnosed
9;22 translocation
in all BM mitoses.
The
karyotype,
determined
from PHA-stim-
peripheral
blast
crisis
blood
been
were made
with Giemsa,
cytogenetic
and
from peripheral
blood cells. When
1 3% of the cells on the cytocen87%
few
were
found
(Fig
iA).
(Fig
and
clear-cut
granulocytic
These
four cells
3A). All the other
Sudan-negative
chromosomes
an
(Fig
extra
1 B, C).
F and
two
They
had
Ph’
chromo-
somes
(Fig 2B). These
cells were anti-GPA-positive
and thus of erythroid
origin
(Fig 1 D). The conclusion
is that
the
erythroid
blast
crisis
affected
only
the
cells
Case 2.
A 48-year-old
woman
had been diagnosed
as having
CML
in chronic
phase.
The karyotype
of
BM
cells
normal.
from
her
was 46,XX,Ph’.
After
BM
contained
The
three
20%
blasts
As
All
constitutional
years,
karyotype
a specimen
aspirated
and
promyelo-
20%
cells
ma’
studied
banding
(68%)
mitotic
Giemsa
cells
Os.
#{149}1
negative
or did not contain
tion had 47 chromosomes
other
slides,
with
the
CALL
21
22
Fig 2.
Conventionally
trypsin
Giemsa-banded
partial
karyotypes from cases 1 and 2 showing
only chromosomes
9. 21 . and
22. (A) Case 1 had in addition
trisomy
for chromosome
19. the
complete
karyotype
being
48.XX.
+ 1 9. + 22q - .t(922).
(B) The
karyotype
of case 2 was 47.XX.
+21 .t(922).
karyo-
black
contained
clear-cut
as well
lymphoid
cell
I antiserum
B, 29
granulo-
cytic granulation
or were Sudan-positive.
had 47 chromosomes.
Those
mitoses
that
These
cells
were Sudan-
myeloid
granula(Fig
1 E). On the
population
comprised
identified
less than
1% of all
the cells,
and there
were
no mitoses
in CALL-Ipositive
cells. The erythroid
population,
as identified
with anti-GPA,
made up 1 5% of the cells on the slides.
On
the anti-GPA-stained
slides,
60 mitoses
ied. Only
two of these
60 mitoses
positive.
The
quality
of these
two
determination
were
stud-
were
anti-GPAmitoses
did not
of chromosome
number.
In con-
clusion,
the mitotically
active
cells in the bone
were
nonlymphoid
and,
with
the exception
marrow
of 2/60
cells,
charac-
nonerythroid.
teristics
cated
ity
The
of the
that
(68%)
or
perhaps
chromosome
karyotype
Thus,
the
staining
of these
cells.
mdi-
a minorof less
All
identified
mitoses
Ph’
and
an extra
G chromosome,
ie,
associated
with
blastic
transformation.
blast
crisis
affected
the myeloid
or
the
monocytic
be
while
cells
the
and
could
mitoses
origin,
myeloid
monocytic
chromosomes
myeloid
cytochemical
majority
they were of myeloid
could
be either
(32%)
had
the
lineages.
DISCUSSION
ln
attempts
cells
whose
how
to
spread
the
structures
here
goals.
better
the near
intact
cell
while
allowing
result
will
origin.
of the
study,
within
Direct
origin
of mitoses.79
the mitotic
spreads,
allows
lineage
the
cell.
approaches
at
experimentation,
be produced
Stenman
and
attempted
to indicate
before.
Earlier
the erythroid
et al’ devised
staining
nucleoplasmic
cells, the authors
also
mosome
identification.
in
to the identification
antigens
of metaphase
cells.
While
the
investigation
was to perform
microfluorimetry
mosomal
method
achieving
When
iron granules
were
the mitosis
was considered
immunofluorescence
is
the
possible
The
aimed
of empiric
undoubtedly
issue
and
the best
the
mitotic
key
membrane
is a compromise
It is the
methods
future.
the
is under
of chromosomes
described
both
and
to identify
karyotype
preserve
cytoplasmic
erythroid
9
the
or Sudan
counted
of mitotic
cells have been
studies
used
iron staining
2$
conventional
revealed
2B). Cytocentrifuge
slides
for simultaneous
cytogenetic
BM
morphologic
analysis.
On slides stained
with
of 43
by the
technique
(Fig
the
of the
lineage.
was
and
whose
48
BM
type 47,XX,+2i,t(9;22)
were made
from
maturation
had the karyotype
46,XX,Ph’
40 mitoses
were in nongranular
cells
with
granulation
the
Giemsa
permit
morphologic
trifuge
slides showed
granulation.
The remaining
of cells
were
mainly
nongranular
blasts
with
lymphocytes.
Four of a total of 44 mitoses
studied
to have
All
trypsin
antigens
seen on
to be of
a method
of
that
acid-labile
aim
in
of their
of chro-
metaphase
reported
having
attempted
chroThe chromosomes
of Raji cells
From www.bloodjournal.org by guest on June 16, 2017. For personal use only.
1 120
TEERENHOVI
itiis
mu
1311 1 LI
#{149}16
UI
.
L
#{149}
A
nil
(3
1 3U
Ex
zaa,
(A) Karyotype
S
$.I
BtIIZI
Fig 3.
a
I has
&aO%
11
,,
.
AeA
of the meyloid
cell shown
in Fig 1A: 46.XX.Ph’.
(B) Karyotype
of the cell shown
$
in Fig 1B: 48.XX.+F.+Ph’.Ph’.
ET
AL
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SIMULTANEOUS
STUDY
OF KARYOTYPE
AND
MORPHOLOGY
Table
1121
1 . Methods
Used
to Id entity
Mitotic
Cells
Chromosome
Method
Cell Lineage
Giemsa
Myeloid
cells,
of other
Sudan black B
Identified
tentative
Staining
identification
Comment
Giemsa
Routine
cells
screening,
As above
Giemsa
Chromosome
heavily
a-Naphthyl
acetate
Monocytes
Hematoxylin
cult
Erythroid
cells
0-banding
The
TRITC
+
possible
identification
positive
Chromosome
esterase
Anti-glycophormn-A
Q-banding
after destaining
identification
because
two
plied
difficult
in
cells
often
of weak
staining
duff
i-
staining
methods
can
be ap-
simultaneously
Anti-Iysozyme
Monocytes
0-banding
As above
TRITC
Anti-kappa/lambda
B lymphocytes
0-banding
As above
+
+ TRITC
could
easily
be
counted,
did
not
allow
applied
specific
chromosomes.
stimulated
by the work
al,2
A method
similar
who,
in eight
but
that
after
activators,
the
mosomes
difference
study
et al.
was
of surface
1gM
chromosome.
stimulation
majority
ours
but
conjunction
methods
identification
of
in part
to ours was used by Bernheim
et
CML
patients
out of 18 found
with
of B cell
were analyzable
between
our
et al is that
fluorescent
banding
The present
ofStenman
evidence
of the occurrence
cells having
the Philadelphia
cluded
the
unequivocal
were
method
on mitotic
They con-
polyclonal
mitoses
B cell
whose
chro-
Ph’ positive.
The major
and that of Bernheim
enables
one to use
also
histochemical
not only
staining
immunofor the
these
logic
and
cytic
leukemia,
mality
in
mitoses
and
t(l5;17)
the
proportion
translocation
mia.’#{176}
As regards
is well
chromosome
suited
in
of
in acute
banding,
conjunction
techniques.
acceptable
However,
Q-bands.
QM
with
immunofluorescence
mitoses
with
promyelocytic
fluorescence
is also
by
produces
a need
banding
methods
that can be used simultaneously
with
hematologic
stains.
Even when
banding
is successful,
one
major
problem
chromosomes
bands
is
are
therefore
aberrations,
change
banded
is well
with
the
present
method
is that
mostly
short
and the number
of
limited.
For this
reason,
small
ie, those
in which
there
is little
-.
dromes,
abnormal
in which
karyotype
about
half
in their
of the patients
BM cells but
have
mostly
are
synan
in
It
with
would
be
of
an abnormality
thrombocythemia’3
of chromosome
16 and
and
of
an abnor-
eosinophilia,’4
and
eosinophils,
respec-
respectively).
Another
interesting
application
study
which
cell lineages
bear the
cells
cells
crisis
in CML.
In case
of this method
is to
karyotypic
markers
I, only
the
had the karyotype
stigmata
of blast
ofgranulocytic
origin had the karyotype
CML
in chronic
common
stem
phase.
This
may
cell being
disease,
mean
have
the
abnormal
erythroid
crisis.
The
typical
of
that
only
the
into cells expressing
by the blast crisis,
the
unaffected.
affected
lymph
only a minority
of cells of Reed-Sternberg
type, most of the cells being lymphocytes.
tion of this method
would
reveal
which
karyotype.
Finally,
nodes
contain
or Hodgkin
The applicacells actually
when
the
pres-
ent method
has been further
improved
to the stage of
detecting
small deletions
and translocations,
it should
be possible
obvious
in chromosome
size, will not be found even in
mitoses.
On the other hand, the present
method
suited
to reveal
numerical
abnormalities
or
distinctive
markers
such as 22q - or Sq
The potential
applications
of the present
method
numerous.
Examples
include
the myelodysplastic
an association
3 and
In Hodgkin’s
for
of
leu-
the erythroid
and
to have morpho-
alterations.’2
cells capable
of differentiating
erythroid
markers
were affected
identification
It usually
there
the
leuke-
20%
tively,
have been found.
Our method
may allow
us to
determine
whether
the observed
chromosomal
abnormalities
occur
in the affected
cell lineages
(ie, mega-
of blast
from
the same
of promyelocytes
About
nonlymphocytic
preleukemic
phase,
lineages
also appear
quantitative
chromosome
Berger
specimen
different
slides
the proportion
metaphases.”
in acute
interest
to know whether
these lineages
have an abnormal karyotype.
In a relatively
small number
of patients
with a myelodysplastic
syndrome
or acute nonlympho-
karyocytes
al’#{176}
used
for counting
normal
terminate
kemia.”
In the
megakaryocytic
recognition
of cell lineage
and chromosome
staining
methods
other
than Q-banding.
Others
have tried
to
identify
mitotic
cells by indirect
methods.
For instance,
et
with
patients
to decide
whether
or not
all leukemic
cells
have a chromosomal
abnormality.
There
are now many
new monoclonal
antibodies
that
identify
myeloid
and lymphoid
cells at various
stages
of differentiation.
When
improved,
the new
cytogenetic
method
described
here should
allow
the
unequivocal
identification
of the cell lineage
and,
perhaps,
mosome
stage of differentiation
abnormality.
It will
that carries
be interesting
the chroto test the
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TEERENHOVI
1 122
extent
to which
the various
specifically
bound
to cells
monoclonal
antibodies
are
with
a given
karyotypic
abnormality.
.
Stenman
spread
of unfixed
staining
2.
5,
Rosenqvist
M,
metaphase
of nuclear
chromosomes
Exp Cell
antigens.
Bernheim
A, Berger
R: Philadelphia
chronic
leukemia.
3.
Dacie
Churchill,
4.
and
JV,
Lewis
1975,
Ranki
26:632,
6.
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1969
1975
iL,
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5, Bussel
B lymphocytes
Res 5:331,
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LC,
Caspersson
T,
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P: Identification
cytocentrifuged
in Viciafava
Gahmberg
CG,
surface
marker
Int J Cancer
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E: Chemical
16:185,
Blackstock
cells
Brown
on
I :844,
A
WM,
Baikie
AG,
bone-marrow
in
Wil-
chronic
1963
method
for
in chromosome
the
positive
identification
preparations
of bone
of
marrow.
Br
1969
AM,
Berger
Bernard
Garson
in acute
OM:
Direct
myeloblastic
R, Bernheim
t(l
et non promyelocytaire.
mia:
London,
A,
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evidence
for involvement
leukaemia.
Lancet
Chromosomes
Daniel
MT,
Valensi
2:1178,
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Nouv
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T
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DAG,
Bennett
iM,
Gralnick
L,
Modest
differentiation
metaphase
Teerenhovi
of early
23:717,
L,
erythroid
P:
Foley
fluorescent
GE,
Cell
Wagh
U,
alkylating
Res 58:128,
Sultan
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DL,
8:181,
Cancer
G,
promyelocytaire
1979
on Chromosomes
Genet
D,
Daniel
C: Proposals
in Leuke-
Cytogenet
2:108,
MT.
HM,
and
of chromosome
Flandrin
Rowley
51 :189,
iD,
Galton
3. Cancer
iM:
associated
Genet
of the
1982
Vardiman
thrombocythemia
No.
G,
for the classification
Br i Haematol
Golomb
leukemia
abnormality
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Exp
I 3. Sweet
myelogenous
differen-
1979
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Vuopio
Catovsky
HR.
myelodysplastic
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leukemia.
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studies
I 980
from
in acute
cells
1 1 . Second
1981
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iM:
erythropoietic
10.
Labaume
leukemia.
of erythroid
PA,
1974
immunofluorescence
1976
5. Andersson
Simonsson
and
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A, T#{246}tterman TH,
Glycophorin
tiation
Practical
Preparation
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Leukemia
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Immunol
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R, Preud’homme
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NR:
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1984 64: 1116-1122
A method for simultaneous study of the karyotype, morphology, and
immunologic phenotype of mitotic cells in hematologic malignancies
L Teerenhovi, S Knuutila, M Ekblom, L Rossi, GH Borgstrom, JK Tallman, L Andersson and A de la
Chapelle
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