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(CANCER
RESEARCH
37. 3228-3237,
September 1977]
Light-Microscopic Morphology of Cell Types Cultured during
Preneoplasia from Foreign Body-reactive Tissues and Films1
Kenneth H. Johnson, Lance C. Buoen, Inge Brand, and K. Gerhard Brand
Department of Veterinary Pathobiology,
College of Veterinary Medicine, University of Minnesota, St. Paul 55108 ¡K.H. J.¡,and Department
Medical School, University of Minnesota, Minneapolis, Minnesota 55455 [L. C. B., I. B., K. G. B.¡
SUMMARY
Cells isolated in vitro from preneoplastic foreign body
(FB)-reactive capsule tissue or surfaces of FB segments
from mice were studied and found to conform to one of four
cell-type categories on the basis of light-microscopic mor
phology, pattern of in vitro appearance, in vitro topographi
cal relationships, and certain karyotype similarities. Euploid
type I (macrophage-like) and II (fibroblast-like) cells pre
dominated in primary cultures and early passages (pas
sages 1 and 2) of cells derived from FB-reactive capsule
tissue. The observation of small numbers of type III cells
(unidentified cell type with unknown karyotype characteris
tics) in passages 1 and 2 of cells from FB-reactive capsule
origin coincided with the deterioration of euploid type II cell
populations and preceded the observation of type IV (endothelial-like) cells. Type IV cells had a pronounced growth
advantage over cell types I, II, and III, resulting in cultures
composed only of type IV cells after three passages. Cul
tures derived from cells attached to the surfaces of FB
segments also conformed to the criteria established for type
IV cells. Of the four cell types identified in this study, type IV
cells were determined to have special importance regarding
the nature of the progenitor cell in FB tumorigenesis, in that
they were aneuploid and eventually produced homologous
sarcomas when injected as a suspension into compatible
hybrid recipient mice. These findings are consistent with
our earlier reported hypothesis implicating certain cells of
the microvasculature as the likely progenitor cells from
which FB sarcomas are derived.
INTRODUCTION
of Microbiology,
and electron microscopy have elucidated morphological
features which suggest that the progenitor cell is a multipotential mesenchymal cell type derived from the local micro
vasculature (14).
More recent investigations in our laboratory have concen
trated on the in vitro isolation and study of FB-induced
capsule- and/or film-attached cells from film-capsule com
plexes removed at various stages postimplantation (8). The
potential usefulness of these in vitro techniques for obtain
ing further information regarding the nature of the progeni
tor cell and various aspects of neoplastic transformation
appears promising because of the preliminary findings that:
(a) initial cultures consisting predominantly of euploid fibroblast- and macrophage-like cells were often gradually
outgrown by morphologically distinct cells with specific
aneuploid karyotypes which were identical with, or closely
related to, tumors derived from corresponding portions of
Tilm-capsule complex left in mice; (b) the cultured aneu
ploid cells frequently gave rise to homologous sarcomas
when implanted in hybrid recipient mice; and (c) the neo
plastic cell determinants were stable in vitro as they were in
vivo, but preneoplastic cell maturation was arrested during
culture, regardless of the number of passages in vitro. The
latter studies have thus demonstrated that preneoplastic
cells from which sarcomas arise many months later can be
isolated and expended in vitro. This provides the opportu
nity to obtain preneoplastic cell preparations from FB-reac
tive tissue and films for analytical studies at defined stages
of preneoplastic maturation.
The primary objective of this investigation was to charac
terize and identify morphologically the various cell types
isolated in vitro from preneoplastic FB-reactive tissues and
films.
The induction of sarcomas in mice by s.c. implantation of
various FB2 materials has been used in our laboratory as a
model to elucidate preneoplastic events concerning, espe
cially, the origin and identification of the progenitor cells
and the nature of neoplastic transformation. These studies
have provided information regarding the appearance time
and location of preneoplastic clones of cells in FB-reactive
tissue and on implant surfaces (6, 7). Studies of FB-induced
sarcomas utilizing special histological staining techniques
1 Supported by USPHS Grant CA 10712 from the National Cancer Institute.
2 The abbreviations
used are: FB, foreign body; MGG. May-GrunwaldGiemsa.
Received January 3. 1977; accepted June 13, 1977.
3228
MATERIALS
AND METHODS
Primary cultures or various passages of 5 different cell
cultures isolated in vitro from preneoplastic capsules or
implant surfaces were utilized for this light-microscopic
study (Table 1). Five mice (CBA/J, CBA/H, and C57BL6/J x
C57BL6/J-bgJbg' F, hybrid) were implanted s.c. in the left
flank area with either single hydrophilic, 15- x 22-mm Millipore filters (0.025-/nm pore size), manufactured by Millipore
Filter Corp., Bedford, Mass., or single 15- x 22- x 0.2-mm
unplasticized vinyl chloride-vinyl acetate copolymer films
(Busse, Great Neck, N. Y.). The basic procedures used for
CANCER
RESEARCH
VOL. 37
Downloaded from cancerres.aacrjournals.org on June 16, 2017. © 1977 American Association for Cancer Research.
Cell Types Cultured in Preneoplastic FB Tumorigenesis
Table 1
Procedural details and derivation of cell cultures used for in vitro light-microscopic studies
Original donor mouse data
Implant data
Cell culture data
Culture
vivo
identifica
source of
tionACulturepno.PrimaryPassage
assage
cellsCapsuleCapsule
implantation
time
code
(mos.)555 no.90229022
(mm)15
xO.215
x22
B
1
Passage 2
Passage 3
Passage 4
Passage 5In
x22
Capsule
VCVA
15 x 22
15 x22
Capsule
VCVA
15 x 22
Capsule
VCVA
15 x 22
CapsuleMaterialVCVA"VCVA
VCVASize
Passage 2
Film
CDEFPrimaryPassage
VCVA
xO.2
xO.2
xO.2
x 0.2
xO.2Total
15 x 22 x 0.2
5
5
5Mouse
strain(C57BL6/J
x C57BL6/Jbgjbgj) normal F, hybrid
Normal F, hybrid
Normal F, hybrid
9022
M
Normal F, hybrid
9022
M
Normal F, hybrid
9022
M
9022SexMM
MMouseNormal F, hybrid
9022
M
(C57BL6/J x C57BL6/Jbg-'bg-') normal F, hybrid
2215
x
2Passage
x2215
2Passage
x2215
BA/ H
x 22 x 0.21.51210e9097910090946129FFFMCBA/JCBA/JCBA/JC
6CapsuleCapsuleCapsuleFilmMF"MFMFVCVA15
" CVA, unplasticized
vinyl chloride-vinyl
acetate copolymer films.
b MF, Millipore filter (0.025-/im pore size).
r Film/capsule
complex transferred in situ after 3 months implantation
in Mouse 6129 to a CBA/H-T6 Mouse (6521); after 4 months in
Mouse 6521, film and attached cells were transferred to a (C57BL10/ScSn
x CBA/H-T6) F, hybrid mouse (6941) for an additional 3
months. Cultured cells (cell line F) were of original donor mouse genotype (6129).
FB implantation are the same as those detailed in our earlier
publications (5, 6).
Preneoplastic FB segments or FB-reactive capsule tissue
for culture were excised at 1, 1.5, 2, 5, and 10 months after
implantation. Details of procedures used for surgical exci
sion of FB segments or FB-reactive tissue for culture or
transfer during preneoplasia are the same as those previ
ously reported (5, 6).
For cell culture (8), preneoplastic, FB-reactive capsule
tissue was minced, treated for 30 min with 0.3% collagenase
at 37°on a magnetic stirrer, and washed with balanced salt
solution. These cells were seeded into 35-mm plastic cul
ture dishes (with an 11- x 22-mm glass coverslip on the floor
of each dish) containing McCoy's 5A modified medium with
antibiotics and 20% newborn calf serum. Cell-laden film
implants were placed directly into 35-mm culture dishes
containing the culture medium. The cultures were passed
routinely (after trypsinization) as soon as monolayers
reached confluence (i.e., after approximately 4 to 6 days for
capsule-derived primary cultures and 3 to 5 weeks for filmattached primary cultures).
Each culture was expanded into 2 culture dishes. Glass
coverslips (with attached cells) on the floors of the culture
dishes were removed from the cultures for morphological
and karyological studies when the cultures had reached
monolayer confluence. The coverslip from 1 of the paired
culture dishes was scored and divided so that one-third was
placed in methanol fixative for subsequent staining with
MGG for light microscopy, and two-thirds were placed in
3% buffered glutaraldehyde for electron microscopy. Cells
attached to the coverslip in the 2nd culture dish were uti
lized for karyological studies (5).
SEPTEMBER 1977
RESULTS
Cell Culture A (primary culture and passages 1 to 5),
derived from preneoplastic capsule tissue enveloping a
plastic film implanted for 5 months, was used as the proto
type for identification and morphological characterization
of FB-reactive cell types and cell populations present in
vitro at various stages of culture (Table 1). Primary cultures
or selected passages of 5 other cell cultures (Cultures B to
F), derived either from FB-reactive capsule tissue or filmattached cells, were studied for comparison and confirma
tion of observations made with Cell Culture A.
Four morphologically distinguishable categories of cell
types, arbitrarily identified by Roman numerals I to IV, were
isolated in vitro from preneoplastic FB-reactive tissues or
FB segments (Table 2). All cells identified and placed in a
particular category had a number of common morphologi
cal features which were demonstrably different from other
categories of cells (Table 2).
Morphology, Time of in Vitro Appearance, in Vitro Topo
graphical Relationships, and Karyotype Characteristics of
Cell Types Isolated from FB-reactive Tissues or Films
Cell Type I: Macrophage-like
Cells
Morphology. Type I cells (Figs. 1 and 2) were predomi
nantly round to spindle or fusiform in shape. Stellate (tripo
lar) shapes were less commonly observed. The terminal
extremities of relatively elongated bipolar type I cells tended
to fan out, forming a fishtail appearance. The cell margins
at these fanned-out extremities were indistinct.
Type I cells had either 1 or 2 nuclei surrounded by a
3229
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K. H. Johnson
et al.
Table 2
Morphology, in vitro topographical relationships,
culture from time
FB-induced
of in vitro
preneoplastic
appearance,capsules
karyotype,
andand
filmstumorigenicity of cell types isolated in
Time of in vitro appearance
of cell population
Morphological characteristics
FB-revitro topo
FB-immic mar
graphical rela active cap
ginsDistinctCytoplasmic
modificationsMany
tionshipsAttachedtissuePR
KaryotypeNot
plant surface
sule
(3-8)"
basoto
to
observed
philic cytospindleCytoplascally
lo
film or sur P-1 (0-3)
plasmic
cated ;
face of type P-2 (0-3)From
dense hetgranulesNucleiAsymmetri
II cellsFrom
erochromatinIn
Cell
Type1Cell
shapeRound
Irregularly
stellate to
fusiform
Indistinct
feathered
edges
None
Smooth contour;. pink
matrix; 1-8
chromacenters
Overlapped
cells
and
crisscross
growth pattern
III
Elongated
fusiform;
few stellate
Distinct
Cytoplasmic
blebs
on
surface of
occasional
cells
Symmetrically
lo
cated; margination of
heterochromatin
IV
Polygonal
to stellate
Relatively
distinct
Many
cytoplasmic
blebs
on
cells at in
termediate
stages of
passage
Many nuclear
blebs; nu
merous
chromacenters; vi
olet-purple
matrix
" Numbers
in parentheses,
percentage;
PR, primary;
PR (92-97)
P-1 (95)
P-2 (87-98)
genicityNT»
EuploidTumori
Not observed
Euploid
NT
Located
on P-1 (2)
film surface P-2 (2-10)
close
to
type II cells
or on sur
face
of
Type
II
cells;
formed
syncytiallike
ar
range
ments
Not observed
Not yet established
NT
Formed pave- P-3 (100)
ment-like
P-4 (100)
P-5 (100)
monolayer
with little or
no
cell
overlap
P-2 (100)
P-6 (100)
Aneuploid
P, passage.
6 NT, In vivo tumorigenicity not tested.
c Cells injected s.c. as a suspension into compatible hybrid recipient mice.
smooth, contoured nuclear membrane. Nuclei were charac
teristically at least slightly eccentrically
placed within the
cells, even in round cell shapes. The nuclei were character
ized by the presence of uniformly dense heterochromatin
that made it impossible to delineate the presence of nu
cleoli.
The cytoplasm of type I cells was uniformly and relatively
intensely basophilic.
Numerous
basophilic
Cytoplasmic
granules (Fig. 2) and occasional vacuoles were characteris
tically present, especially
concentrated
in juxtanuclear
areas.
Time of in Vitro Appearance.
Type I cells were observed
as 1 of the 2 cell types predominating
in confluent monolayers of primary cultures of preneoplastic FB-reactive capsule
tissue (Cultures A and C) that had been removed after 1.5
(Culture C) and 5 (Culture A) months of implantation. Type I
cells represented 8% (Culture A) and 3% (Culture C) of the
cells in these primary cultures. This cell type represented
3% of the cells from passage 1 of Culture A but was not
observed in confluent monolayers of any of the subsequent
3230
passages studied from Culture A (passages 2 to 5) and was
not observed in a confluent monolayer of passage 2 of
Culture D. Three % of the cells from the confluent monolayer stage of passage 2 of Culture E were identified as type
I cells.
Type I cells were not observed in confluent monolayers of
passage 2 of Culture B or passage 6 of Culture F, both
derived from film-attached
cells. [Primary cultures of filmattached cells were not studied in these experiments,
but
we have demonstrated
the presence of macrophage-type
cells on implant surfaces in many previous experiments (7,
13).]
In Vitro Topographical Relationships. Type I cells were
either attached directly to the coverslip surface or were
located on the surface of thin cytoplasmic extensions of
type II cells also present in primary cultures and early pas
sages. Direct cell-to-cell contact between type I cells was
rarely seen, even when they were present in colonies or
foci.
Chromosomes. Type I cells were invariably euploid (i.e.,
CANCER
RESEARCH
VOL. 37
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Cell Types Cultured in Preneoplastic FB Tumorigenesis
a less proliferative but functionally active stage. If type III
and/or IV cells were present in these early stages of growth
in vitro, they were obscured by the initial rapid proliferation
of fibroblast-like type II cells. Aneuploid type IV cells were
demonstrable when the growth of type II cells subsided.
Passage at this point resulted in the appearance of signifi
cant numbers of type IV cells approximately midway
through the passage (e.g., at 5 to 6 days of passage 3 of
Culture A) and a pure population of aneuploid type IV cells
at the stage of monolayer confluency (e.g., at 11 days of
passage 3 of Culture A).
The possible relationship of type III cells to aneuploid type
IV cells is not clearly understood on the basis of these lightmicroscopic studies alone. The observation of type III cells
in culture coincided with the deterioration of euploid fibro
blast-like populations (cell type II) and the 1st appearance of
type IV cells. The disappearance of type III cells coincided
with the progressive and rapid expansion of aneuploid type
IV cells. Insufficient numbers of type III cells in mitosis
precluded any conclusions regarding the karyotype charac
teristics of the cell type.
On the basis of the observations of this study, it is possi
ble to propose at least 3 hypotheses that could account for
the changing predominance of cell types: (a) small numbers
of preneoplastic type IV cells were present in primary cul
tures but were obscured during this period by the rapid
proliferation of fibroblast-like cells (i.e., type II cells); (b)
type III cells, present in small numbers in early cultures,
represent the progenitor cells from which type IV aneuploid
cells develop under selective and preferential conditions
provided in the in vitro environment; and (c) type IV cells,
although recognized in the study to be distinct from type II
cells on the basis of light-microscopic morphology and
growth characteristics, represent in vitro aneuploid variants
of the earlier-appearing, euploid, fibroblast-like cells (type II
cells). Further information concerning the possible relation
ships and distinctions between cell types II, III, and IV is
provided by our electron-microscopic studies of these cell
types (to be reported in a subsequent paper).
As was discussed earlier, all cells in the 2 cultures derived
from cells attached to the surfaces of FB implants (i.e.,
passage 2 of Culture B and passage 6 of Culture F) con
formed to the criteria established for type IV cells. These
observations possibly indicate that the surface of FB im
plants in these cases provided the same preferential stimu
lus for development and growth of type IV cells before
culture (i.e., in vivo) as the cell cultures provided for these
cells in vitro. Consequently, type IV cells already present on
the FB surface at the time of excision apparently had an
early growth advantage in vitro (e.g., passage 2 of Culture
B) over other cell types also possibly present on the FB
segments.
In that the nature and origin of cell types cannot be
determined with absolute certainty by their light-micro
scopic morphology in culture, the 4 cell types observed in
this in vitro study were arbitrarily identified by Roman nu
merals to avoid confusion that would result from designa
tion of specific names without further ultrastructural and
histochemical conformation. However, on the basis of lightmicroscopic studies of MGG-stained preparations, certain
SEPTEMBER
1977
morphological and growth characteristics of the cell types
observed in this study could be correlated with in vitro
features reported for several specific cell types.
The size (15- to 50-jiim diameter), shape (round to spin
dle), and presence of numerous cytoplasmic granules
and/or vacuoles characteristic of type I cells are consistent
with functionally active macrophages.
Type II cells, which tended to grow in overlapping and
crisscross arrays, were observed as large (up to 400 /¿m
long) stellate to fusiform cells with indistinct feathered
edges, numerous fibrillar cytoplasmic components, and
closely associated extracellular deposits of collagen-like
material. These morphological and growth characteristics
are generally consistent with those described for fibroblasts
or fibroblast-like cells in vitro. These characteristics of type
II cells also conform in general to those described for
smooth muscle cells in vitro, but type II cells did not form
distinct bands of parallel cells which are characteristic of
smooth muscle cells (10). The presence of collagen forma
tion is not adequate evidence to distinguish between fibro
blasts and smooth muscle cells in vitro because smooth
muscle cells (like fibroblasts) have been reported to secrete
collagen (15, 17). It is thus apparent that an accurate mor
phological distinction between fibroblasts and smooth
muscle cells in vitro must be made with the electron micro
scope (17).
Type III cells characteristically were very slender (approxi
mately 10 /xm in diameter), bipolar cells up to 200 ¡¿m
long
that often formed linear cords of overlapping cells. These
cells had a single symmetrically located nucleus with heav
ily marginated heterochromatin. Stellate type III cells, with 3
or 4 finely tapered cytoplasmic processes that provided
points of cell-to-cell contact, were also apparent. Attempts
to specifically identify type III cells based on these lightmicroscopic studies alone are admittedly speculative, but
these cells do appear to have some features in common
with certain cells isolated in vitro from retinal capillaries.
Retinal capillaries present in expiants of retina cultured in
vitro produce cord-like outgrowths of cells (1) that some
what resemble the overlapping cell networks observed in
type III cells in culture. These cells, derived from retinal
capillaries and identified as primitive vascular mesenchymal
cells, were demonstrated to have the capacity to differen
tiate into endothelial cells and intramural pericytes (1).
Type IV cells were polygonal to stellate and, in contrast to
type II cells, had relatively short cytoplasmic processes,
more clearly defined cell margins, and a more homogenous
cytoplasm with less fibrillar banding. Type IV cells were also
generally more uniform in size and shape as compared to
type II cells and often contained cytoplasmic vacuoles pre
sumed to be lipid. The growth characteristics of type IV cells
were significantly different from type II cells in that the
former cells grew to confluence, forming a monolayer with
little or no evidence of piling up or cell overlap. These
morphological features and the tendency to form monolayers of uniformly spaced cells in pavement-like or mosaic
patterns are consistent with the characteristics reported for
in vitro cultured endothelial cells (2, 11, 12, 18, 19).
Cytoplasmic blebs were commonly observed with type IV
cells in intermediate stages of passage (passages 3 and 4) of
3233
Downloaded from cancerres.aacrjournals.org on June 16, 2017. © 1977 American Association for Cancer Research.
K. H. Johnson
et al.
Culture A and only occasionally with type III cells. Cytoplasmic blebs were not present in later passages of Culture A,
nor were they present in cultures of type IV cells obtained
from film-attached cells (passage 2 of Culture B and pas
sage 6 of Culture F). In that type IV cells were demonstrated
in this and earlier studies to be aneuploid cells capable of
producing tumors when injected into compatible recipient
mice, it is possible that the cytoplasmic blebs associated
with these cells are linked to a phase of neoplastic transfor
mation. Cytoplasmic blebs were not associated with the
euploid cell types that predominated in the primary cultures
and early passages of Culture A. The possibility that cyto
plasmic blebbing is linked to the process of neoplastic
transformation is further supported by scanning electronmicroscopic studies of in vitro transformed BALB/3T3
mouse embryo cells (16). In the latter studies, SV40, murine
sarcoma virus, and spontaneous transformants of the
BALB/3T3 cell line were shown to have numerous 1.0 to 5.0/nm cytoplasmic blebs not present in the parent cell line.
Also in the context of the present study, BALB/3T3 cells,
which were originally considered to be fibroblast cells, are
now considered to have morphological and growth charac
teristics that are more consistent with cells derived from
small blood vessels (i.e., endothelial cells or pericytes) (9,
16). The morphological and growth characteristics reported
for BALB/3T3 cells are in fact not unlike those of the type IV
cells identified in the present study.
Recent experiments have further demonstrated that
BALB/3T3 cells, which cannot divide in vitro unless at
tached to a solid substrate and are nontumorigenic when
inoculated s.c. into syngeneic mice, produce tumors (ma
lignant hemangioendotheliomas) when they are inoculated
s.c. after being attached to glass beads (3) or plastic films
(4). These findings, which demonstrate that BALB/3T3 cells
are tumorigenic in animals when attached to a solid sub
strate, provide an additional link of similarity between
BALB/3T3 cells and our in vitro isolated type IV cells. He
mangioendotheliomas or hemangiosarcomas are in fact va
rieties of sarcomas induced by the experimental implanta
tion of plastic or glass films (14).
The findings of this in vitro study are considered to be
consistent with our earlier reported hypothesis (14) impli
cating cells of the local microvasculature as the likely pro
genitor or parent cells from which FB sarcomas are derived.
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1973.
Fig. 1. Primary culture of FB-reactive tissue origin. The monolayer is composed predominantly of type II cells and a few type I cells (arrows). MGG, x 256.
Fig. 2. Focus of type I cells (round, spindle, and stellate forms) from a primary culture of FB-reactive tissue. The eccentrically placed nuclei contain
uniformly dense heterochromatin. Numerous basophilic cytoplasmic granules are evident. MGG, x 1024.
Fig. 3. Large stellate type II cell from a primary culture of FB-reactive tissue. The smooth-contoured nucleus contains 2 nucleoli and multiple, smaller
chromacenters. Faintly visible fibrillar components (arrows) are evident in the less basophilic cytoplasmic matrix. MGG, x 1024.
Fig. 4. Dense culture (passage 1 of FB-reactive tissue origin) composed predominantly of overlapping and unidirectional type II cells with early appearance
of a few typical fusiform type III cells (arrows). MGG. x 256.
Fig. 5. High-magnification photomicrograph of 2 fusiform type III cells present in Fig. 4. These cells have hyperchromatic nuclei that are symmetrically
located with the cell cytoplasm. The cytoplasm of type III cells is more intensely basophilic than the cytoplasm of the underlying type II cells. MGG, x 640.
Fig. 6. Two closely associated type III cells (passage 1 of FB-reactive tissue origin) located on the surface of thin cytoplasmic extensions of large underlying
type II cells. A few clear vacuoles (arrows) are evident in the cytoplasm of the type III cells. MGG, x 1024.
Fig. 7. Nonconfluent monolayer of polygonal type IV cells (passage 3 of FB-reactive tissue origin). These cells have more well-defined cell margins and
more homogeneously basophilic cytoplasm than type II cells observed in earlier passages. A large number of intensely basophilic blebs (arrow) are evident on
the cytoplasmic membrane of 1 type IV cell. MGG. x 384.
Fig. 8. High-magnification photomicrograph of 2 type IV cells (passage 4 of FB-reactive tissue origin). Irregularities in nuclear membrane contour are
manifested by areas of slight indentation or evagination to form nuclear blebs. The cytoplasm is characterized by numerous clear vacuoles, distinct margins,
and basophilic blebs (arrows) concentrated on the extremities of pseudopodia. MGG. x 640.
Fig. 9. Confluent monolayer composed of uniformly spaced type IV cells (passage 5 of FB-reactive tissue origin). There is little or no evidence of poling up
or cell overlap. A significant degree of irregularity in nuclear membrane contour is apparent, and cytoplasmic lipid vacuoles are apparent in some type IV cells.
MGG, x 205.
Fig. 10. High-magnification photomicrograph of uniformly spaced type IV cells (passage 5 of FB-reactive tissue origin) without directional orientation.
MGG, x 512.
3234
CANCER
RESEARCH
VOL. 37
Downloaded from cancerres.aacrjournals.org on June 16, 2017. © 1977 American Association for Cancer Research.
Cell Types Cultured
in Preneoplastic
FB Jumorigenesis
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3235
Downloaded from cancerres.aacrjournals.org on June 16, 2017. © 1977 American Association for Cancer Research.
K H. Johnson
3236
et al.
CANCER
RESEARCH
Downloaded from cancerres.aacrjournals.org on June 16, 2017. © 1977 American Association for Cancer Research.
VOL. 37
Cell Types Cultured
in Preneoplastic
FB Tumorigenesis
r
SEPTEMBER
1977
Downloaded from cancerres.aacrjournals.org on June 16, 2017. © 1977 American Association for Cancer Research.
3237
Light-Microscopic Morphology of Cell Types Cultured during
Preneoplasia from Foreign Body-reactive Tissues and Films
Kenneth H. Johnson, Lance C. Buoen, Inge Brand, et al.
Cancer Res 1977;37:3228-3237.
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