Tissue Culture Model of Transitional Cell ... - Cancer Research

Tissue Culture Model of Transitional Cell Carcinoma:
Characterization of Twenty-two Human Urothelial Cell Lines
John R. W. Masters, Peter J. Hepburn, Lawrence Walker, et al.
Cancer Res 1986;46:3630-3636.
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[CANCER RESEARCH 46, 3630-3636, July 1986]
Tissue Culture Model of Transitional Cell Carcinoma: Characterization of
Twenty-two Human Urothelial Cell Lines
John R. W. Masters,1 Peter J. Hepburn, Lawrence Walker, Wilma J. Highman, Ludwik K. Trejdosiewicz,
Susan Povey, Mohamed Parkar, Bridget T. Hill, Peter R. Riddle, and Leonard M. Franks
Department ofHistopathology, St. Paul's Hospital, Institute of Urology, 24 Endell Street, London WC2H 9AE, United Kingdom [P. J. H., J. R. W. M., L. W., W. J. H.,
L. K. T.]; Gallon Laboratory, M. R. C. Human Biochemical Genetics Unit, University College, 4 Stephenson Way, London NWl 2HE, United Kingdom [S. P., M. P.];
and Imperial Cancer Research Fund, Lincoln's Inn Fields, London WC2A 3PX, United Kingdom [B. T. H., P. R. R., L. M. F.J
ABSTRACT
Twenty-two continuous cell lines derived from normal and neoplastic
urothelium, maintained under identical culture conditions, were charac
terized in terms of isozyme phenotype, tumorigenicity, and xenograft
morphology following xenotransplantation to nude mice, cylological ap
pearance, in vitro growth rate, labelling index, and colony-forming effi
ciency, in parallel with separate studies of in vitro drug sensitivities and
monoclonal antibody reactivities. Three groups were identified: (a) dis
tinct lines with differing isozyme patterns, a broad spectrum of growth
characteristics, and xenograft morphologies similar to the histopathology
of the parent tumors after periods of up to 17 yr following establishment
in vitro; (b) cross-contaminated sublines (maintained separately in differ
ent laboratories for periods of up to 10 yr), with identical isozyme patterns
and similar growth characteristics, but differing markedly in tumorigenic
ity and xenograft morphology; and (c) lines derived from normal uro
thelium which were nontumorigenic and had an isozyme pattern usually
only encountered in untransformed cells. These data indicate that cell
lines representative of human transitional cell carcinomas can be selected
on the basis of xenograft morphology and isozyme patterns, and that a
panel of lines derived from normal and neoplastic urothelium could
provide a model system to study the biology and treatment of this disease.
INTRODUCTION
The natural history and therapy of human tumors are dictated
primarily by the site and histopathology of the disease. Never
theless, among tumors of any one histológica! type there is a
broad spectrum of biological behavior and response to particu
lar treatment regimen. Consequently model systems are needed
which accurately represent not only individual tumors, but also
reflect the range of properties of each histológica! type of
disease. One model system that might encompass these dual
criteria is a panel of continuous cell lines derived from tumors
of one histological type. In this study we investigated the extent
to which these two criteria are fulfilled by a panel of cell lines
derived from TCC2 of the human bladder. No previous attempt
has been made systematically to examine under standardized
conditions the properties of a large series of human TCC cell
lines, although there are numerous reports concerning individ
ual or small numbers of lines ( 1-4).
MATERIALS AND METHODS
Details concerning the origin of the lines used in this study are shown
in Table 1. Eighteen lines were derived from transitional cell carcino
mas, one from a squamous cell carcinoma, and three from normal
urothelium. With the exceptions of 253J, VM-CUB-I, and VM-CUB-
III (obtained from Dr. J. Fogh, Sloan-Kettering Institute for Cancer
Research, Rye, NY), all the lines were obtained either from the origi
nator or laboratory of origin: RT4 and RT112 (this laboratory);
Received 7/2/85; revised 12/27/85; accepted 3/17/86.
1To whom requests for reprints should be addressed.
2The abbreviations used are: TCC, transitional cell carcinoma; ADA, adenosine
deaminase; CFE, colony-forming efficiency; LI, tritiated iIn mulino labeling
index; PBSA, calcium- and magnesium-free phosphate-buffered saline; PDT,
population doubling time; PPLO, pleuropneumonia-like organism.
3630
COLO232 (Professor G. E. Moore, Denver General Hospital, Denver,
CO); HT1197 and HT1376 (Professor S. Rasheed, University of South
ern California, Los Angeles, CA); HU456, HU609, and HU961T (Dr.
M. Vilien, Fibiger Laboratory, Copenhagen, Denmark); MGH-U1 and
MGH-U2 (Professor G. R. Prout, Massachusetts General Hospital,
Boston, MA); PS1 (Dr. E. J. Sanford, Pennsylvania State University,
State College, PA); KK47 (Professor H. I lisa/unii. Kanazawa Univer
sity, Kanazawa, Japan); HS0767 (Dr. W. A. Nelson-Rees, Naval Biosciences
Laboratory, Berkeley, CA); TCCSUP, SCaBER, J82COT, and
T24 (Dr. C. O'Toole, Cambridge, United Kingdom); and HCV29 (Dr.
J. Fogh).
Cell Culture. All lines were grown routinely on plastic as monolayers
in 25-cm2 flasks (Nunc; Gibco, Paisley, Scotland), in RPMI 1640
medium (Gibco), supplemented with 5% heat-inactivated fetal bovine
serum (Flow Laboratories, Irvine, Scotland) derived from a single batch,
and 2 HIML-glutamine (Gibco) at 36.5°Cin a humidified atmosphere
of 5% CO2 in air. Each cell line was used over a restricted number of
passages (maximum of 10), using stocks held in liquid nitrogen (see
Table 1). For subculturing, monolayers were detached using 0.01%
irypsin (Difco Laboratories, London, England; Difco 1:250) plus
0.003% versene (BDH Chemicals, Poole, England; EDTA disodium
salt) in PBSA.
PPLO Screening. PPLO contamination was screened by staining
with acetoorcein (5) and by inoculation of cells together with their
culture medium into PPLO agar on a feeder layer of BHK21/13C cells,
as previously described (6).
Isozyme Analysis. A maximum of 13 polymorphic enzymes was
isozyme typed in each cell line. Exponentially growing cells were
detached, washed once in medium and once in PBSA, centrifugea at
250 x g for 10 min between each step, transferred in PBSA in 1-ml
aliquots containing approximately 10 cells to freezing vials (Nunc),
and again centrifuged at 250 x g for 10 min. The PBSA was decanted,
and the cells were left to air dry for 15 min before storage in liquid
nitrogen until analysis. The enzymes were typed by horizontal starch
gel electrophoresis as previously described (7), except for a-fucosidase
which was examined by isoelectric focusing (8).
Tumorigenicity in Nude Mice. Exponentially growing cells were de
tached and washed twice in serum and glutamine-free (unsupplemented)
medium. Approximately 10 cells in 0.1 ml of unsupplemented medium
were injected s.c. into the flank of a nude mouse (nu/nu), using at least
four replicates for each cell line. The mice were examined weekly for
gross evidence of tumor development and killed by cervical dislocation
when tumors had grown to 0.5-1.0 cm in diameter. The tumors were
excised, fixed in Baker's formol calcium, and processed using routine
histological procedures. Paraffin sections were stained with hematoxylin:eosin.
Cytology. Air-dried cell smears were stained with May-Grunwald/
Giemsa (R. A. Lamb, BDH Chemicals, London, England), and cell
smears fixed in 95% alcohol were Papanicolaou stained (Ortho Diag
nostics, High Wycombe, England).
Colony-forming Efficiency on Plastic. Exponentially growing cells
were detached enzymatically, and single-cell suspensions were prepared
by repeated passage through a 19 gauge needle. Serial 50% dilutions
were plated in triplicate in 5-cm plastic Petri dishes (Nunc) to yield
viable cell numbers ranging from 50-6400/dish. The cells were incu
bated for between 8 and 28 days, depending on the growth rate of the
cell line. Cell lines requiring an incubation period of 14 days or more
to develop colonies were medium-changed at 7-day intervals. The
colonies were fixed in methanol (BDH Chemicals; Analar grade) and
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CHARACTERIZATION OF HUMAN UROTHELIAL CELL LINES
Cell line
of tumor
Table 1 Origin of cell lines
stage
r culture of
prior
designationT24MGH-U1 nos.51-60103-112129-13854-6387-9634-4367-7682-9136-4535-4435-4459-6814-2316-2549-5831-40132-14178-8723-3211-2018-27Origin
biopsyRecurrence diseaseNR°T2
of
gradeG,G,NRG,G,G,G,G4G.G,G,G.NRNRG,G)NRY
established1970197219751975NR1972196719721972197319731974197519751975
patientFMMMMMMMMFFFMMMMMMNRMFPatient's
therapyNoneNRNRNoneNone4200
bladderBladder in
(EJ)MGH-U2HU456HU961TJ82COTRT4253JHT1197HT
primaryBladder
primaryBladder
primaryBladder
primaryBladder
primaryRecurrence
bladderRetroperitoneallymph
in
node metas
tasisRecurrence
bladderBladder in
1376RT112TCCSUPVM-CUB-IVM-CUB-IIIPS1COLO
primaryBladder
primaryBladder
primaryBladder
primaryBladder
primaryBladder
primaryBladder
232KK47SCaBERHCV29HS0767HU609Passage primaryBladder
primanBladder
primarysquamous
cell car
cinomaNormal
frompatient bladder
with blad
cancerNormal der
frompatient bladder
with pros
cancerNormal tate
frompatient ureter
renalcellwith
cancerClinical
" NR, not recorded.
* C. C. Rigby, personal communication.
minimumNRTiT,T,TjT«T;
stained with 10% Giemsa (Gurrs Improved R66; BDH Chemicals).
Colonies containing more than 50 cells were scored using a binocular
dissecting microscope, and the mean CFE derived from a minimum of
three separate experiments was calculated.
CFE =
no. of colonies
no. of viable cells seeded
x 100
Population Doubling Times. Exponentially growing cells were plated
in 3.5-cm dishes (Nunc) containing 5 ml of medium, at numbers ranging
in serial 50% dilutions from 768,000-24,000 cells/dish, with 2 repli
cates for each cell concentration at each time point. After 24, 48, 72,
and 96 h of incubation the cells were enzymically detached, and a single
cell suspension was produced by repeated passage through a 19 gauge
needle. Cells were suspended in a known volume of filtered PBSA, and
the numbers in two replicate 0.5-ml aliquots of each sample were
determined using a Coulter Counter (Coulter Electronics, Luton, Eng
land). The mean results from a minimum of two experiments were
plotted on semilogarithmic graph paper, and the PDT was calculated
for cell concentrations growing exponentially between 48 and 96 h after
initial plating, using the formula PDT = In2/ [In (Nt/No)]~', where / is
time interval (48 h), Nt is number of cells at 96 h, and No is number of
cells at 48 h.
Labeling Indices. Exponentially growing cells were plated in 5-cm
plastic Petri dishes containing 5 ml of medium. Following a 48-h
incubation period to facilitate the resumption of an exponential growth
rate, the medium was discarded, and fresh medium containing [methyl-
3H]thymidine (20 ¿iCi/ml;Amersham International, Amersham, Eng
land; 25 Ci/mmol) was added to each dish. After a 15-min pulse, the
dishes were rinsed 3 times with unsupplemented medium at 4°Cfor 5
min each, fixed by three 5-min washes in 95% ethanol at 4"C, and left
inverted to air dry. The fixed cells were coated with chrome alum/
gelatin (9) and prepared for autoradiography as described previously
(10). Briefly, the cells were covered with a layer of Ilford K5 emulsion
(Ilford Nuclear Research, Knutsford, England) diluted 1:3 with distilled
water. Following a 7-day exposure at 4°C,the emulsion was developed
minimumT2
minimumNRT4NRNRT,T»NRTjHistológica!
preop-erativelyGold
rads
yrearlierNRNoneNoneNoneNoneNRNRNone4000
grains 2
mopreoperativelyNRNoneNRNRRef.404142434344
rads 2
using Kodak D19 (Kodak-Pathé,Chalon-Sur-Saone, France) diluted
1:1 with distilled water. A minimum of 500 nuclei was counted, and
those covered by more than 15 individual silver grains were scored as
positive. Data are derived from two separate experiments.
Time-lapse Cinemicroscopy. The apparatus used has been described
(11). Five-cm Petri dishes containing cells were transferred to an
Olympus IMT inverted microscope fitted with a Perspex incubator,
Bolex H16J camera, and Olympus control unit. Kodak 7454 or Kodak
Infocapture AHU 1454 microfilm was used, and the negatives were
analyzed on a LW analyzing projector.
RESULTS
Data from 22 cell lines are described. Five of the lines (PS1,
COLO232, J82, KK47, and J82 COT) were PPLO positive on
repeated examination and were examined in less detail in order
to avoid contamination.
Isozyme Analysis. Among the 22 cell lines, there were 16 that
differed at one or more alíeles(Table 2), indicating each is
unique in origin. In contrast, T24 and five other lines (J82,
MGH-U1, MGH-U2, HU456, HU961T) were identical at each
locus, indicating that these almost certainly have been crosscontaminated.
Original stocks of J82, designated J82 COT,
differ from T24 and have an isozyme pattern identical to that
of another cell line derived from the same patient, providing
further evidence that the subline J82 has been cross-contami
nated with T24 in some stocks. Two further lines, VM-CUB-I
and VM-CUB-III, had identical isozyme phenotypes, suggest
ing cross-contamination. None of the cell lines had the same
isozyme pattern as HeLa cells.
The three lines derived from normal urothelium (HCV29,
HS0767, HU609) all had an ADA isozyme of slow electrophoretic
mobility (designated "tissue" in Table 2). This phenotype
is common in untransformed cells in culture and, among the
3631
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Cell line
designation
CHARACTERIZATION OF HUMAN UROTHELIAL CELL LINES
Table 2 Isozyme profiles of the bladder cell lines
Enzyme loci
PGM1" PGM3 GOTM COTS ESD AK1
ADA PGD G6PD GLO FUCA PGP ACPI
T24MGH-UI
(EJ)MGH-U2HU456HU961TJ82J82COTRT1I2
;ABI:ABI:ABI:ABI:ABI;A
B i
0RT4
2-1TCCSUP
:2-1
]2I2-112-1
2-1MD
:A
A2-1A B
B
B
2
2253J
1HTII97
1HT
2VM-CUB-I 1376
2-1VM-CUB-I11
2-1KK47
2-1'issue
2)
1A
A
2A ND
1A
1A
B
ND
B
B
B
B
J-1.-1
BABABABMD
1COL0232
2-1PS1
2A
:A
A
B 2 1MD BMD
1SCaBER
2-1HCV29
2-1HU609
1HSO767
2-122
1ND
1A
1fissue
;[issili- A
IPissue A
1.-1MD11MDMDMD 1VDMDMDMD :.-i.-i:!-l:!-l:!-ll!!M!-l'AAC
B
A
ND
B
B
ND
2-1
1
2
ND
11!-l BMD
BMD
1MDMDBABABABABABACABNDABABA-1
A1
CAVD
1;_;.¡_)_;_!-l!-l_ B
* PGM l and 3, first and third loci of phosphoglucomutase; GOTM and S, mitochondria! and soluble glutamate-oxaloacetate transaminase; ESD, esterase D; AKI.
adcnylatc kinase; PGD, 6-phosphogluconate dehydrogenase; G6PD, glucose-6-phosphate dehydrogenase; GLO, glyoxalase: FUCA, a-fucosidase; PGP, phosphoglycollaie
phosphatase; ACPI, first locus of acid phosphatase; ND. not done: Tissue, not typable because ADA complexed with glycoprotein.
tumor-derived lines, was only observed in HT1197.
Tumorigenicity. Details concerning the growth of the cell
lines as xenografts are summarized in Table 3. Two tumorderived
lines (T24 and TCCSUP) and the three lines derived
from normal urothelium (HCV29, HU609, HS0767) failed to
develop tumors during a maximum follow-up period of 2 yr.
Independent morphological descriptions of each tumor by
two experienced urological histopathologists were in agreement
(summarized in Table 3). Replicate xenografts derived from the
same cell line had similar morphologies. The tumors produced
by RT4 and RT112 were morphologically similar to those of
the original biopsies taken in 1967 and 1973, respectively (Figs.
1 to 4). Areas of squamous metaplasia were more common in
the xenografts, perhaps reflecting differences in the hormonal
environment in the mice. HT 1376 and HT 1197 produced
poorly differentiated tumors similar in microscopic appearance
to those illustrated in the original description of these lines
(12). Thus, in the cases in which direct comparison could be
mo)"
tumorigenicityCell
line
designationT24MGH-UI
appear
cellsDifferentiated
ance of cultured
Table 3 Cell line morphology and
TCCDifferentiated
tumorsGj, not form
(EJ)MGH-U2HU456HU961TRT112RT4253JHTI376HT1197VM-CUB-11ITCCSUPCytological
TCCDifferentiated
MR*Gj, possibly TCC with spindle cell areas and high
TCCDifferentiated
MR< possibly TCC with spindle cell areas and high
TCCDifferentiated
metaplasia( . , TCC with squamous
TCCDifferentiated
metaplasiaGi.2,
' •

Fig. 1. Parent tumor of RT112. Papillary transitional cell carcinoma of the
human bladder. H & E, x 240.
CHARACTERIZATION OF HUMAN UROTHELIAL CELL LINES
F'8- 3- Parent tumor of RT4- Papillary transitional cell carcinoma of the
h""""1 bladder. H & E, x 240.
i*.l::v»-?;^'íV:r^:*•;...-.;.,¿. ? 5; ^,-f; r ;-'. „.-•v*v»>»v .
88ÄIP Ìli; IfÜä
Fig. 2. RT1I2 xenograft showing similar morphology to that of the parent Fig. 4. RT4 xenograft. showing similar morphology to that of the parent
tumor in Fig. 1. H & E, x 240. tumor in Fig. 3. H & E, x 240.
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CHARACTERIZATION OF HUMAN UROTHELIAL CELL LINES
Growth Characteristics. The in vitro growth characteristics of Xenotransplantation has been used to demonstrate that most
the cell lines are listed in Table 4. The similarities in these
features between T24 and its cross-contaminated sublines con
human tumor cell lines retain a neoplastic phenotype and
produce xenografts with a morphology compatible with that of
trast with the wide variation among the other distinct lines. the tumor of origin (21, 22). In the largest series of cell lines
There is some association between a long PDT and both low studied, 127 of 162 (78%) produced tumors, and in every case
CFE and LI. An exception is HT1197 which, despite having the histopathology correlated with that of the tumor of origin
the longest PDT and lowest CFE, has a relatively high LI of
37.3%. This observation was investigated further using time-
(23). In this and earlier studies of both human (2, 3) and rat
(24) bladder tumor cell lines, the ability to form tumors with
lapse cinemicroscopy to compare the cell division times and morphologies similar to those of the tumors of origin was
cell loss rates of HT1197 and T24 (see Table 5). The lack of retained.
association between the PDT and LI of HT1197 may be ex Tumorigenicity was highly variable among the five T24 sub-
plained by the high death rate of HT1197 (35%) compared with lines tested. Cloning of one of these sublines, MGH-U1 (des
that of T24 (0.5%).
ignated EJ in some studies), also resulted in lines differing
widely in tumorigenicity (25). Loss of tumorigenicity has also
been observed in melanoma (26) and other cell lines (23). These
DISCUSSION
differences may reflect changes in the immunogenicity of the
This paper describes certain biological characteristics of a
panel of cell lines derived from TCC of the human bladder,
maintained under identical and standardized culture conditions.
In parallel studies, we measured in vitro drug sensitivities (13)
and reactivities with a panel of monoclonal antibodies derived
from one of these cell lines (14). These data provide a basis on
which to evaluate a panel of cell lines as a model system for
tumors of this histológica! type.
Isozyme analysis can be used to identify cross-contamination
between cell lines of the same or another species and, if tissue
from the patient is available, establish that the cells are derived
from that individual (15). Isozyme phenotypes are retained in
vitro and usually remain stable over many years in culture (16).
In this study, 16 lines were shown to have distinct isozyme
phenotypes, indicating derivation from different individuals,
while 6 lines appeared to be cross-contaminated. Bladder tumor
cell lines appear to have been cross-contaminated with T24
cells in at least 3 separate laboratories. The isozyme patterns
of some of these lines have been described previously (15, 17-
20), and our data are in agreement.
cells, perhaps reflected by the changes we observed in the T24
sublines in cell surface antigen expression demonstrated with
monoclonal antibodies (14). T24 and its sublines could provide
a model system for studying certain of the factors involved in
tumorigenicity, and they are of particular interest as T24 was
the first human cell line shown to contain a transfectable
oncogene (27).
The in vitro growth characteristics of the T24 sublines were
similar, contrasting with the wide variation among the distinct
cell lines. The relative stability and reproducibility of these data
probably result from the use of standardized conditions. All the
lines were grown in one type of tissue culture medium, supple
mented with a single batch of fetal bovine serum over a re
stricted passage range. Alteration in any one of these conditions
can radically alter the properties of cells in vitro, and it is
essential that comparisons between cell lines are made using
standardized culture conditions. The growth rates of all of these
lines are considerably higher than those observed in human
bladder tumors, either as measured directly in patients (28) or
indirectly in biopsies in vitro (29). Similarly, the colony-forming
efficiencies on plastic (ranging from 3-87%) were far higher
Table 4 Growth characteristics of cell lines
than those achieved by cells cultured
human bladder tumor biopsies (30).
in agar directly from
Cell line Population dou-
(h)MGH-U2MGH-U1
designation bling time
efficiency on
In addition to characterizing
plastic (%)877657618416222752743Labeling
index535254515129423927321237
preliminary studies were made
cell lines derived from TCC,
on three lines derived from
normal urothelium. In support
(EJ)HU961TT24HU456RT1I2VM-CUB-3TCCSUP253JHT1376RT4HT1197192021212124262828313761Colony-forming
tissue, all three lines expressed
of their origin from normal
the untransformed phenotype
of the ADA enzyme, in contrast to only 1 of 13 of the tumorderived
lines. The untransformed phenotype is seen in virtually
all normal adult fibroblast cultures, but it is rare in transformed
cells in culture (31). However, the ability of cells to grow
indefinitely in vitro is usually regarded as an abnormal feature,
and therefore the relevance of these lines as a model for normal
human urothelium is uncertain. Normal urothelium can be
cultured directly from rat (32, 33) and human (34) bladders,
Table 5 Estimates of cell loss rate and intermitotic times ofHT1197 and T24 lines followed for two cell generations by time-lapse cinemicroscopy
Some cells were lost from the field of view, as tabulated, either as a result of migration or detachment from the substrate.
Cell lossTotal
cellsLost
viewDiedIntermitotic
from field of
(h)Total time
cellsRangeMeanGeneration
15812211827.8-63.54S.6HT1197Generation
2397141323.6-102.248.6Total971935
(36%)3123.6-102.246.9Generation
3634
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Copyright © 1986 American Association for Cancer Research
I77207513.4-26.417.1T24Generation
215216113512.3-20.916.4Total229181
(0.5%)21012.3-26.416.6

and it is now possible to propagate large numbers of human
normal urothelial cells in defined medium (35). Spontaneous
transformation in such cultures has been observed with rat, but
not human, urothelium (36).
The ultimate goal of these studies is to assess the value of
these cell lines as a model for TCC. Cell lines can be selected
which are representative of their tumors of origin on the basis
of isozyme analysis and xenotransplantation. Nevertheless,
some characteristics, such as growth rate and clonogenicity,
alter as a result of in vitro culture, and these changes must be
taken into account when using cell lines as a model system.
The panel of cell lines shows a wide range of morphology,
degree of histological differentiation, biochemical properties,
growth characteristics, sensitivities to chemotherapeutic drugs
(13), and reactivities with monoclonal antibodies (14). Similar
heterogeneity has been observed in cell lines derived from
human lung tumors (37), ovarian carcinomas (38), and mela
nomas (26). These data indicate that a panel of cell lines could
be selected reflecting the spectrum of properties exhibited by
each histological type of tumor. For use as a model system, it
is also essential that the properties of the cell lines remain
relatively stable in long-term culture. However, tumors are
inherently heterogeneous and capable of generating diversity,
for instance by developing drug-resistant phenotypes (39). Sta
bility in long-term culture was investigated using the T24 sublines,
which had been maintained in various laboratories under
different culture conditions, separately for periods of up to 10
yr. The sublines were similar in their isozyme patterns, growth
rates, and in vitro drug sensitivities, but markedly different in
tumorigenicity and reactivity with certain monoclonal antibod
ies (14). To limit such differences and ensure comparability
between and within laboratories, it will be necessary to use a
common stock of each cell line over a limited number of
passages and under identical culture conditions.
In conclusion, these data support the concept that a panel of
cell lines derived from human TCC could provide a model
system for this disease, but also illustrate some of the inherent
problems. Disadvantages include the need for rigorous stand
ardization of culture conditions, the requirement to characterize
large numbers of cell lines and exclude cross-contamination
and PPLO infection, and the fact that certain characteristics
are not stable in long-term culture. Advantages include the
unlimited supply of cells derived from each human tumor and
the stability of features such as isozyme phenotypes, in vitro
drug sensitivities, and the ability to generate tumors histopathologically
similar to the tumor of origin.
ACKNOWLEDGMENTS
K. A. Wallace and D. E. Bennett carried out the xenografting
procedure in the Animal House Unit at the Imperial Cancer Research
Fund Laboratories. We are indebted to Dr. M. C. Parkinson and Dr.
K. M. Cameron for their support and advice.
REFERENCES
1. Hepburn, P. J., and Masters, J. R. W. The biological characteristics of
continuous cell lines derived from human bladder. In: G. T. Bryan and S. M.
Cohen (eds.). The Pathology of Bladder Cancer. Vol. 2, pp. 213-227. Boca
Raton, FL: CRC Press, Inc., 1983.
2. Grossman, H. B., Wedemeyer, G.. and Ren, L. UM-UC-1 and UM-UC-2:
characterization of two new human transitional cell carcinoma lines. J. I ml..
132: 834-837, 1984.
3. Kyriazis, A. A., Kyriazis. A. P.. McCombs, W. B.. and Peterson, W. D.
Morphological, biological, and biochemical characteristics of human bladder
transitional cell carcinomas grown in tissue culture and in nude mice. Cancer
Res.. 44:3997-4005, 1984.
CHARACTERIZATION OF HUMAN UROTHELIAL CELL LINES
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