From www.bloodjournal.org by guest on June 17, 2017. For personal use only.
Use
Cell
of Spleen
Organ
Regeneration
Cultures
Following
By Harald
After
lethal
levels
exceeded
The
irradiation
T
24
factor
HE
hr
and
initiating
regeneration
between
and
various
are
interest
of the
tissues
indicated
the
operation
control
processes
while
other in vivo
of considbut
have
complexity
formation
stem cells
of local
Cells
0.5 ml
EBSS
was
cellularity
were
undertaken
simple
factors
on
the
system
and avoid
secondary
to endotoxemia.
The system
observed
in organ
repopulation
in
organ
an
culture
spleens
MATERIALS
cells
were
such
for
mice
mice.
Groups
of 4 irradiated,
irradiated
Bone
organ
cell
forced
wk
12-14
tissue.”
dishes
containing
supplemented
(Upjohn,
in each
strips
of 0.45s
shown
in Fig.
For every
irradiated
cells
some
24 hr
nonirradiated
adjustment.
be
of
each
mm
Cu and
conditions.
the
The
Preparation
Femoral
flushing
Blood.
focal
dose
old,
were
Vol.
56.
body
kV,
distance
was
developed
used
to
throughout
irradiation
I 5 mA.
was
Fifteen
to ten slices
from
different
containing
pooled
with
Eisen’s
5 (November),
that
described
spleens
were
placed
as from
only
were
regions
9 normal
anatomical
fragments
were
mice.
or spleens
different
In
from
were
cut
micrometric
regions
filter
spleen
were
bone
All spleens
0.02%
as
that
syngeneic
slicer with
of each
was
media,
fragments.
9 mice
tissue
containing
size
with
organ
mice
used.
sponge
7 mm
l0
as well
Medium
on the sponge,
from
irradiated
from
x
saturated
with
Petri
surgical
top of the Millipore
on the
the organ
60
removed
mice
Eagle’s
for the
transplanted
fetal
plastic
of
paper.
were
harvested
EDTA.
The
Five
in I
culture
at 37#{176}C
in an
incubated
of 10% CO2.
After
from
were
as a supp,t
from
x
was
served
solution
trypsin
atmosphere
a 20
Gelfoam
the
to 20 slices
placed
the
2, 4, and 6 days of incubation,
3 different
spleens
from
the
filter
3 cultures
were
harvested
paper
into
slices
lute
filter
with
full
a Philips
and
used was 0.2
of
the
Cancer
Medical
by
containing
slices
gently
scraping
trypsin
solution.
All
mm at 37#{176}C.Single
the trypsinized
slices
cell
twice
a 0.25%
for 30
washing
backscatter
tal.
cold
1980
from
20 normal
Balanced
Salt
mice
Solution
in part
by
Hall
P.O.
© 1980
the
and
April
reprint
Institute
Carden
Md.).
Bonn,
29. /980;
accepted
requests
to Dr.
ofMedical
H.
the
M.
Research
Ca-25972
supported
by
Germany.
July
Research.
Anti-Cancer
Grants
was
Insti-
Victoria.
Medical
Institute
v.
Hall
of the
and
West
H.
Eliza
Hospital,
Fellowship
Cancer
e. V. 5300
and
Health
National
Victoria,
by Grune
Walter
Melbourne
National
(Bethesda.
Krebshilfe
3050,
Unit,
Royal
by the
Victoria.
Submitted
Address
Research
Research.
CA-22492
Deutsche
Eliza
cells were
No.
using
The
50 cm
marrow
shaft
and
and
with
using a mechanical
were
I 27 rad/min.
Suspensions
bone
as
sieve
METHODS
of Cell
bone
each
skin
rate
prepared
in 85-mm
Mich,)
previously,
spleen
dishes
due
was
prepared
Modified
and transplanted
slices
Canberra,
total
trans-
steel
lymph
A Gelfoam
spleens
Council.
at 250
and
essentially
calfserum.
and
experiments
were
membrane
experiment,
lethally
was
Dulbecco’s
Millipore
of
rad
in
pipetting.
of
After
I . These
Supported
800
cells
20 normal
were
fetal
20%
dish.
Council
operating
l0
and
a stainless
of thymus,’
cultures
Kalamazoo,
placed
used
culture
I 5 ml
with
Irradiation
given
suspensions
through
system
organ
Organ
spleen
fragments
AND
pipetting,
by gentle
in EBSS
culture
Australia.
were
a
transplanted,
and
untreated
normal
dislocation
and individual
femurs
and
marrow
were
for the
experiments.
machine
as
Cultures
ml of 0.25%
as those
analysis
initially
and
by gentle
dispersed
From
Mice
of spleen
marrow
20 of 40 irradiated
Spleens
Organ
Mice
x-ray
the
cell
marrow.
cultures
from
into
were
trypsinized
suspensions were prepared
after
mice,
in the
dispersed
removed.
into 0.2-mm
microenvironmental
effects
chosen
raft
technique,
organ
cultures.9
female
progenitor
were
planted,
nonirradiated
and
mice were killed
by cervical
organ
C57B1
and
incomplete
injected
nonirradiated
The
system that
would
sustain
hemopoietic
regeneration
following
irradiation
and marrow
transplantation,
simple enough
to permit
analysis
of the influence
the Gelfoam
support
thymus
spleen
previously
changes
observed
are
but follow the devel-
a relatively
humoral
total
remained
continuous
marrow
of endotoxemia.78
present
experiments
added
cells,
microen-
early in the regenerative
studies
have implicated
seems
apparent
that some of the
not direct
responses
to irradiation
to develop
in the
described.
the operation
of factors
able to initiate
stem cells into
active
cell cycle.3’4
Studies
using
in vitro hemopoietic
colonies
have also indicated
perturbations
in GM-CSF
levels, controlling
granulocyte
and macrophage
proliferation
following
irradiation.5’6
In the latter
case,
it
attempt
marrow
Transplantation
E. Mandel
regeneration
(EBSS).
body
in vivo.
have
opment
The
10
but
excluding
controlling
studies
on hemopoietic
colony
and marrow
by transplanted
vironmental
period,’2
of
Marrow
Thomas
and
spleen,
populations
whole
Extensive
in the spleen
clearly
Metcalf,
injection
in the
transplantation,
following
marrow
transplantation
and
practical
to analyze
because
of interactions
by the
1 2 days
and
Progenitor
regeneration.
MECHANISMS
irradiation
and
erable
theoretical
proved difficult
Donald
of progenitor
irradiation
in splenic
hemopoietic
followed
within
capacity
after
Hemopoietic
Irradiation
von Melchner,
mice
levels
regenerative
prepared
significant
of C57B1
pretreatment
exceptional
slices
to Monitor
23, /980.
von
Melchner,
Royal
Walter
Melbourne
and
Hospi-
Australia.
& Stratton.
Inc.
0006-4971/80/5605-0026$0I.00/0
917
From www.bloodjournal.org by guest on June 17, 2017. For personal use only.
918
VON
irradiation
and
MELCHNER.
marrow
METCALF,
transplantation
slight
increase
in spleen
2 and 5, a major
rise occurred
8 with
the
this
period.
was
observed
cellularity
by day
subnormal
by day
supplemented
in this
agar
with
way
cultures.
were
20%
In addition,
suspensions
were
calf
serum.
and
counts
with
suspensions
cells
smears
stained
were
Cell
for colony-forming
cytocentrifuge
prepared
sa. Differential
fetal
assayed
(CFC)
of the
in
same
cell
cells
under
oil
an
atmosphere
May-Grunwald/Giem-
performed
on
100
in
after
(CFC)
In Vivo
CFCs
2 and
I 2.
In
within
normal
level
cellular-
until
levels
Levels
frequency
time
of
points
day
still
5
then
being
the
spleen,
CFCs/105
normal
frequency.
Although
the
nucleated
irradiation
3. CFCs
in both
and
reappeared
the marrow
was observed
in the bone
plateau
between
days
the
was more rapid,
reaching
a peak
in the Bone
after
is shown
in Fig.
transplantation
and spleen.
A steady
increase
marrow, reaching a subnormal
and
prepared
Cell
different
transplantation
very early
levels
the
marrow
increase
cellularity
After
between
days 5 and
12.
Spleen
change
at
normal
beyond
I 2. In contrast,
no significant
slowly,
with
The
in EBSS
overshoot
ity showed
increased
cells
Fig. 1 . (A) Sen
organ
culture
showing
the G&foam
raft
covered
by two Millipore
strips supporting
the spleen organ slices.
(B) Morphology
of cells harvested
from organ
cultures
of normal
spleens
(upper)
and irradiated
and transplanted
spleens
(lower)
after 6 days in culture.
( x 1000; May-Gr#{252}nwald/Giemsa.)
achieving
A slight
Colony-Forming
Marrow
and
cellularity
between
MANDEL
2).
(Fig.
an intitial
days
AND
increase
in
frequency
5
of
exceeding
normal
levels by day
on day 8 that was 20 times
the
frequency
of
CFCs
in
the
bone
immersion.
Agar
Cultures
Cells
were
containing
cultivated
10%
of agar
The
medium.
0.6%
Bacto
agar
agar
the
for
its
(0.2
source
to
were
cellular
volume
and
40%
with
fetal
as
strength
calf
serum
factor.
This
growth
and
pure
cell
previously
-
of
mixture
double
spleen
prepared
colony
as
200
condi-
,-
described,’2
stimulus
was
used
of granulocyte,
macro-
mixed
as well
erythroid
‘0
IQ
scored
content
placed
on gridded
room
temperature,
stained
with
of
colonies,
every
glass
hydrogen
solution.
Slides
for
using
colonies
fixed
in acetone
160
NORMAL
RANGE
SPLEEN
were
w/v
oil using
added
was
with
a Zeiss
as mean
unless
cell
the
benzidine
and
counter-
Both
counts
Marrow
bone
showed
and
Spleen
80-
40
x 1000
errors
of the
±
otherwise
Cellularity
and
spleen
an initial
fall during
RANGE
_
z_
MARROW
standard
stated.
2
DAYS
marrow
NORMAL
BQE
-
7_
at
content
RESULTS
Bone
SPLEEN
0
was
staining,
to
-
microscope.
values
experiments
at
then
in water.
their
light
and
before
benzidine
solution
to
iust
and
to dry
10 mm
dihydrochloride
acid.
2 mm
according
expressed
separate
for
a 2% safranin
typed
under
were
30%
stained
with
acetic
allowed
for
benzidine
in 0.5%
pipette
120
-I
I.Li
colonies
Pasteur
were
the
determine
sequential
a fine
3
microscope
To
After
were
peroxide
were
dissection
individual
30-45
slides.
solution
I mm
of 4-6
Olympus
magnifications.
dish
slides
magnifications
Results
an
x 35
benzidine/safranin;
0.04%
Colonies
at
from
in a 0.2%
stained
using
lighting
removed
mean
an equal
240
I ml
CFCs.
indirect
used
containing
of Cultures
Cultures
were
mI/dish),
granulocyte-macrophage,
Scoring
dishes
water
Medium
stimulate
as megakaryocyte
with
was
distilled
of colony-stimulating
ability
phage,
37#{176}Cin
Petri
Pokeweed-mitogen-stimulated
medium
was
at
plastic
in
Eagle’s
ea’2
tioned
7 days
medium
(Difco)
Dulbecco’sModified
described
for
in 35-mm
CO2
4
AFTER
6
8
10
12
TRANSPLANTATION
In Vivo
total
the
first
viable
cell
48 hr after
Fig. 2.
Bone marrow
and spleen
cellularity
intervals
after
irradiation
and transplantation.
standard
errors of 4 separate
experiments
each
at different
time
Mean
values
±
using 4 mice.
From www.bloodjournal.org by guest on June 17, 2017. For personal use only.
SPLEEN
ORGAN
919
CULTURES
media
may
lated
toxic
changing
variable
have
the
into
excluded
could
become
products
exhausted
culture
medium
To
introducing
the day 8 time
further
consideration.
No
after
2 days of culturing
nontransplanted
observed
normal
mice
normal
from
spleens
and
no significant
between
the
cellularity
and
mice
the cellularity
transplanted
viable
cells
irradiated
differences
of
of
with
in
Table
time
1 and
periods
Fig.
1
in organ
. The
spleens
spleens
I O bone
cells 24 hr prior to spleen
removal.
The morphology
of the cells recovered
kept for various
avoid
another
point
was
be recovered
were
accumu-
breakdown.
and
the system,
from
or have
of cellular
of
derived
marrow
from
culture
spleens
is shown
lymphoid
content
of
irradiated
transplanted
spleens
was
subnormal
throughout
the culture
period
but after 6 days in organ
culture, an increase in the percentage
of early granulocytic cells was observed.
CFC
Levels
in Spleen
In Fig.
spleens
total CFC
the cultures.
mice
Fig. 3.
Colony-forming
cell frequency
in the
spleen
at different
time intervals
after irradiation
tion. Mean values
± SE of 4 separate
experiments.
bone marrow
and
and transplanta-
numbers
levels
marrow
always
exceeded
that
in the
spleen,
it can
be
calculated
from the data
that
the total
content
of
CFCs
in the spleen
during
the regenerative
period
greatly
exceeded
the levels
in the
normal
spleen,
whereas
the total number
of CFCs
in the marrow
did
not
achieve
normal
levels
during
the observation
pen-
od.
No CFCs
were
spleen
of
during
the time
detected
lethally
in the
irradiated
period
bone
marrow
nontransplanted
covered
by these
experiments.
In groups
of nonirradiated
marrow
cells, no differences
were observed
in the marrow
with normal
controls.
mice receiving
in CFC numbers
or spleen
when
Cellularity
Culture
Table
cells
in Spleen
I presents
recovered
spleens
kept
cell numbers
Organ
the
from
in organ
in normal
absolute
and
mice
numbers
lO bone
and type
compared
of
viable
the normal
and
transplanted
culture.
It can be seen that the
spleens
decreased
continuously
during
the 6 days
of organ
culture,
whereas
in the
spleens
removed
from
irradiated
and
transplanted
mice, cell numbers
remained
relatively
constant
after
an initial
decrease
during
the first 2 days of culture.
Virtually
no cells could be recovered
from either group
on day 8 of organ
culture,
by which
time the culture
rise
total
in organ
culture
content in the same
Spleens
increased
6 days
the
Cultures
the calculated
4A,
kept
Organ
from
number
of CFCs
irradiated
and transplanted
their
CFC
content
seven-fold
of organ
culture
whereas
total
of normal
spleens
The
decreased.
during
CFC
rise
in CFC
in spleen
organ
cultures
closely
paralleled
observed
in vivo in the spleen
of irradiated
planted
mice. However,
the increase
CFC
numbers
was somewhat
lated
in vivo
(Figs.
2 and
3).
in
is compared
with
the
spleens
at the start
of
When
the
trans-
in the total calcuhigher
(nine
fold)
total
calculated
CFC
numbers
in spleens
removed
from
irradiated
and
transplanted
mice
were
compared
directly
with
the
total calculated
CFC numbers
in normal
spleens,
as
shown in Fig. 4B, it can be seen that, despite
variability
between
spleens
tation
fold
individual
removed
exceeded
after
again
spleen
6 days
with
CFC
in organ
the in
numbers
irradiated
and
be 3 times
higher
spleens,
24 hr after
the normal
the
CFC
content
of
irradiation
and transplanspleen
CFC
content
two
culture.
This
correlated
vivo difference
between
normal
and CFC
numbers
of spleens
in
transplanted
6 days
mice,
after
which
were
found
to
transplantation.
In control
organ cultures
of normal
spleens,
the
total number
of CFCs
fell from day 0 to day 4 but did
not fall further
by day 6, despite
the continuing
fall in
total
cellularity
between
spleens
(Table
I ). No significant
differences
CFC
content
of normal
spleens
and
removed
from
nonirradiated
animals
transwith i07 cells 24 hr previously
were
found
the
planted
during the 6 days of organ culture.
Thus,
experiment,
after 6 days of organ culture,
mice
injected
with
iO
marrow
cells
in a typical
spleens
from
contained
an
From www.bloodjournal.org by guest on June 17, 2017. For personal use only.
VON
920
Table 1 . Total
Cellularity
and Differential
Cell Count
of Spleens
Kept
Percent
Days in
Organ
Organ
Normal
Undifter-
Cellularity
Culture
spleen
x
Myeloblasts.
MANDEL
Culture
of
Nucleated
cytes.
Myelocytes
Blasts
AND
Polymorphs
Lympho-
Mono-
Eosino-
cytes
cytes
phils
Red
Blood
Plasma
Cells
Cells
0
0
26.7
±
8.5
1
0
0
92
2
0
5
2
11.3
±
2.4
2
0
0
92
2
0
3
1
4
7.3
±
2.6
1
1
1
64
32
0
1
0
6
4.3
±
1.5
0
2
3
62
29
0
3
1
0
2.9
±
0.9
0
0
14
16
70
0
0
0
transplanted
2
1 .5
±
0.4
0
0
13
12
73
1
0
1
spleen
4
1.5
±
0.3
4
3
3
16
73
1
0
0
6
1.6±0.7
8
11
7
10
63
0
0
1
and
Irradiated
are
Results
1 5 slices
from
average
(see
Table
normal
CFC
exp
as means
ressed
SD from
±
5-6
experiments
each,
of 5.9 ±
1 and
1.8 x 106 cells and 145
Fig. 4B for corresponding
±
25
data
CFC
for
The
mixed
within
different
in the
relative
Bone
Marrow
frequencies
and
of granulocyte
(G-CFC),
FC),
mixed
granulocyte-macropure
and
mixed
erythroid
CFC)
and megakaryocytic
CFCs
(MEG-CFC)
the bone marrow
and spleen
were determined
intervals
Differences
after
mice.
In
of three
spleens.
Total
cell counts
w ere derived
fro m a pool of
pure
percentage
grown
and
recipient
organ-cultured
mixed
erythroid
marrow
and
spleens,
spleen
a
than
slightly
higher
in control
increased
was
larger
Apart
observed
size
from
and
then
these
colonies
those
grown
differences,
spleens
did
from
CFCs
transplanted
DISCUSSION
at
points
it
of cells
was
G-CFCs
mice.
and spleen
of
2, and 5 days
colonies
of
were of generally
the in vivo spleen.
(E-
grafting
and in organ-cultured
between
the relative
frequencies
after transplantation
(Table
2). At these time
was of interest
that the relative
frequency
the
use
the CFC
types grown
from organ-cultured
not appear
to be significantly
different
grown
from the spleen
of irradiated
Spleen
of different
CFC
types in bone marrow
recipient
mice were observed
at 2.5
hr.
forming
the
from
spleens).
Types
spleens.
involving
each spleen.
macrophage
( M-C
phage (GM-CFC),
in
in Organ
METCALF.
Metamyelo-
entiated
1O
MELCHNER.
In confirmation
of earlier
data,’3
these
have shown that the spleen
of irradiated
planted with bone marrow
cells exhibits
hemopoietic
regeneration
than
the bone
least
as assessed
content.
An
by total
essentially
cell regeneration
cultures from such mice,
sponding
Since
assumed
cellularity
similar
was achieved
and
pattern
in isolated
experiments
mice transmore
active
marrow,
at
progenitor
cell
of progenitor
spleen
organ
although
there was no comerise in total cellularity.
seeding
of the injected
marrow
cells can be
to be complete
by 24 hr posttransplanta-
Fig. 4.
Colony-forming
cells recovered
from spleens
kept
in organ
culture.
(#{149}-#{149})
Spleens
from
irradiated
transplanted
mice;
(O-O)
spleens
from
normal
mice.
Mean
values
±
SE of 6 separate
experiments.
Total
colony-forming
cells (B) were calculated
from a pool of 15
slices from each spleen cultured.
From www.bloodjournal.org by guest on June 17, 2017. For personal use only.
SPLEEN
ORGAN
921
CULTURES
Table
2.
Morphology
of Colonies
Grown
After
Time
After
the
Bone
(Days)
Marrow
of 1 O Bone
Days in
Organ
Culture
Transplantation
Cells Cultured
From
Transplantation
and Spleen
Marrow
at Different
Percentage
G
Time
Points
Cells
M
of Colonies
E/Mixed
GM
Meg
0.1
0
10
±
2
65
±
2.5
16
±
1.5
5
±
1.5
0.3
±
0.3
2
0
38
±
6
35
±
2
22
±
3.5
5
±
1
0.3
±
0.3
5
0
30
±
3
32
±
1
32
±
3.5
3
±
0.5
0.6
±
0.5
8
0
13
±
1.5
51
±
2.5
37
±
2
0.6
±
0.3
12
0
26±1
0
2 1
±
2
4 1
±
8
36
±
6
0.5
±
0.5
0.1
0
28
±
3.5
47
±
1
24
±
3.5
0.6
±
0.6
2
0
32±5
Bonemarrow
Normal
control
Spleen
89±1
0
36±2
36±7
1.7±0.8
5
0
19±3
0
15
±
0.4
32
±
3.5
45
±
7
2.2
±
0.3
12
0
20
±
2.5
41
±
9
35
±
4
0.7
±
0.6
Normalcontrol
0
22±6
Normalspleen
37±3
38
5±1.5
27±2
4
1.3
4
±
0.2
±
0.9
9±3
1.5
0.8
1.3
34
20±2
49±4
30±4
0
0
6
43±4
25±2
27±4
0
0
±
1.5
0.6
1.3±0.6
1.3±1.3
1
±
2
±
25
1
4
±
4
1.4
±
0.6±0.5
8
42±6
1
8±2
23±4.5
30±9
0
±
±
1
2
40
±
4
45
±
4
13
±
2
0.3
±
0.2
0.5
±
andtransplanted
1
4
30
±
3
40
±
3
27
±
4
1.4
±
0.6
1.6
±
1
spleen
1
6
25
±
5
37
±
10
35
±
7
0.8
±
0.6
0.6
±
0.6
Irradiated
Results
are expressed
as means
G. granulocytic,
timepoint.
colonies;
Meg,
M,
SE of 4 separate
±
macrophage;
GM.
experiments
mixed
using
cells from
4 mice;
E/mixed,
30-40
sequential
combined
total
colonies
of pure
were
erythroid
sampled
and
at each
mixed
erythroid
megakaryocytic.
tion,’3”4
the similar
regeneration
poietic precursor
cells in isolated
pattern
of hemospleen
organ
cultures
to that
(a)
observed
in vivo
spleen
microenvironment
ing such regeneration,
regeneration
observed
implies
that
the
colonies
whether
isolated
rise
in progenitor
cell
levels
grown
in vitro from
were
equal
in size
is due
regenerating
if not larger
normal
spleen
to CFU-S,’5
suggesting
that
are
Despite
certain
limitations,
the
described
here
using
thin
slices
of
appear
to be a promising
method
for
gating
the mechanisms
controlling
hemopoietic
regeneration
following
marrow
populations.
efficiency
of CFCs
into the
to be I 0 times
lower
when
CFCs
accumulating
in the spleen
originated
from injected
CFCs.
to generation
of progenitor
cells from injected
multipotential
stem cells or to a limited
capacity
for progenitor cell self-replication.
However,
it should
be noted
that colonies
populations
from
the seeding
been
found
compared
must be capable
of regulatand (b) that the marked
splenic
compared
with that occurring
the
grown
Furthermore,
spleen
has
in the marrow
does not necessarily
depend
on extensive seeding
of precursor
cells
from
the marrow
or
elsewhere.
It cannot
be determined
from
the present
experiments
pooled
granulocytic/macrophage;
0.3
few
of
likely
the
to have
organ
cultures
the adult
organ
further
investiearly
events
in
irradiation
and
transplantation.
ACKNOWLEDGMENT
spleen
than
The
authors
are
to Ann
indebted
Williams
for
skillful
technical
assistance.
REFERENCES
I
.
Trentin
tiation
by
stromal
(HIM).
Am
i Pathol
2.
Wolf
hemopoietic
cells.
i Exp
3. Gidali
in partially
4.
Lord
proliferation
226,
5.
colony
ii:
NS,
Determination
ii:
stroma
i, Lajtha
LG:
mice.
BI,
Ki,
Mon
in
stem
cell
6.
differen-
Hemopoietic
colony
studies.
of
V. Effect
pluripotent
of
stem
Kinet
5:147-157,
EG: A stimulator
Wright
regenerating
of haemopoietic
Tissue
bone
marrow.
cell turnover
of stem
Biomedicine
cell
27:223-
1977
Hall
7. Chang
BM:
The
Bri
of
effects
factor
Haematol
and
whole
in vitro
17:553-561,
body
irradiation
on
forming
cells
colony
1969
CF.
stimulating
Soc Exp
Med
Biol
Ryan
M:
serum
in the
P. Bealmear
factor
mice.
Proc
9.
Mandel
thymocytes
10.
Effects
M:
factor
levels
Soc
P. Stohlman
in irradiated
Exp
of
Biol
F, Wilson
germfree
Med
and
140:478-480,
iG,
Mandel
flora
CFW
on
of
levels
mice.
Proc
Levin
i,
infection
levels
in
Sullivan
and
tolerant
R, Bealmear
irradiation
and
on
P.
serum
CF
non-tolerant
1978
Kennedy
and
of microbial
of irradiated
1973
H,
bacterial
factor
TE,
Effects
in serums
144:177-180,
51:229-244,
in vivo
Tew
Pollard
P, Cohen
colony-stimulating
mice. Blood
stimulating
bonemarrow.
CFW
8. Quesenberry
1972
Quesenberry
stimulating
1972
colony
1968
Regulation
AA,
colony
conventional
on differentiation
Cell
Morley
R: Serum
microenvironments
1971
127:205-214,
irradiated
marrow
inductive
65:621-628,
Trentin
organ
Med
of bone
hemopoietic
MM:
in vitro.
TE:
The
differentiation
Immunology
The
maintenance
35:31
of
7-331
and
.
murine
1978
regulation
of
From www.bloodjournal.org by guest on June 17, 2017. For personal use only.
VON
922
serum
antibody
retained
II
.
Teale
lymphocyte
vitro
12.
node
poietic
levels.
in lymphoid
fetal
JM,
Mandel
function
and
organ
Metcalf
cells
culture
D,
stimulating
indicating
J Immunol
TE:
Ontogenetic
susceptibility
GR:
J Exp
a
role
for
120:1063-1069,
tolerance
system.
Johnson
of conditioned
colony
Evidence
follicles.
Med
Production
development
in vivo
by spleen
with
erythroid
activity.
J Cell
Physiol
and
96:3
Metcalf
B-
in an in
14.
and
lymph
other
hemo-
hemopoietic
1978
irradiated
Moore
MAS:
METCALF.
Haemopoietic
AND
Cells.
MANDEL
Amsterdam,
I 971.
Lahiri
of colony
in irradiated
15. von
1980
1-42,
D,
North-Holland,
of
and
151:429-446,
medium
13.
anitgen
1978
MELCHNER.
SK,
forming
mice.
Melchner
Van
Putten
units
from
Cell
precursor
mice.
Leuk
LM:
bone
Distribution
marrow
and
Tissue
Kinet
2:21-28,
H, Metcalf
D: Differential
cells
in
the
spleen
Res October/November,
and
spleen
multiplication
after
1969
seeding
and
bone
1980
injection
of injected
marrow
of
From www.bloodjournal.org by guest on June 17, 2017. For personal use only.
1980 56: 917-922
Use of spleen organ cultures to monitor hemopoietic progenitor cell
regeneration following irradiation and marrow transplantation
H von Melchner, D Metcalf and TE Mandel
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