`
`ANTICANCER RESEARCH 12: 1035-1054 (1992)
`
`:so-1005
`
`Morphological and Immunocytochemical Characteristics of
`
`
`
`
`Human Tumor Cell Lines for Use in a Disease-Oriented
`
`Anticanc�r Drug Screen
`
`SHERMAN F. STINSON1
`, MICHAEL C. ALLEY1
`, WILLIAM C. KOPP2
`, LESLIE A.
`
`, HEINZ-H. FIEBIG3
`
`, SUSAN KENNEY1, JEAN KELLER I and MICHAEL R. BOYD1
`MULLENDORE1
`, ANGELA F. PITTMAN1
`1 Laboratory of Drug Discovery Research and Development, Developmental Therapeutics Program, Division of Cancer
`
`
`
`
`
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`
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`
`
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`Treatment, National Cancer Institute, Building 1052, Room 121, Frederick, MD 21702-1201;
`
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`2Clinical Immu11ology Services, Program Resources, /11c.l DynCorp, Frederick Cancer Research a11d Development Center,
`
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`Frederick, MD21702-1201, U.S.A.;
`
`
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`3 Department of internal Medicine, University of Freiburg, Freiburg, Germany
`
`H (AR) is
`contains
`texes are
`ne.
`ublished
`the legal
`aper has
`
`and jour­
`. Reviews
`
`83, 1984,
`92: lnsti-
`A panel of 60 human tumor cell lines is currently
`features. Most of these lines were poorly differentiated. Lines
`
`
`
`Abstract.
`10.00 per
`
`
`being used in the U.S. National Cancer /nstitute's in vitro
`
`
`
`derived from large-cell and squamous-eel/ cancers also showed
`ther con­
`
`
`
`anticancer drug screen. The panel is organized into 7 sub­
`
`
`
`some characteristics consistent with their reported origins,
`subscrip­
`Personal
`
`
`
`panels; 6 leukemia/lymphoma lines comprise one subpanel,
`
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`except for one line which showed immunocytochemical and
`;$ 300.00
`
`
`and 54 other lines are organized into subpanels representing
`
`
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`morphologic characteristics consistent with rhabdomyosarco­
`tage and
`
`
`
`
`
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`solid tumors of the central nervous system (CNS), colon, lung,
`
`
`ma. The 2 lines derived from small cell lung cancer (SCLC)
`,laced at
`
`
`
`
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`ovaries, kidneys and melanomas. In the present study, the
`
`
`lacked neurosecretory granules and 3 other SCLC markers but
`the pub­
`
`
`
`leukemia and lymphoma cell lines were analyzed by flow
`
`
`showed morhologic features consistent with SCLC. Most
`' to J.G.
`
`
`
`cytometry for appropriate CD antigens; all but 1 line showed
`
`
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`melanoma cell lines strongly expressed melanoma-associated
`:mcer Re­
`, Editorial
`
`
`
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`patterns of expression consistent with their reported deriva­
`
`
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`antigens and were morphologically similar to human melano­
`
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`tions. The solid tumor lines were characterized individually
`
`
`
`ma. Five lines produced premelanosomes, melanosomes or
`requests
`
`
`
`using morphological and immunocytochemical techniques to
`
`
`melanin. Most of the renal cancer cell lines showed morpholo­
`·oduction
`
`
`
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`determine their relative degrees of representativity for the
`
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`gic or immunocytochemical features consistent with renal clear
`
`
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`cell carcinoma. Collectively, these morphological and im­
`
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`
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`subpanels within which they are currently grouped. Histologic­
`
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`munocytochemical analyses provide information concerning
`
`
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`al, histochemical and ultrastructural examinations were per­
`
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`formed on cell lines grown under identical conventional cul­
`
`
`
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`tissue of origin, tumor type, degree of differentiation and other
`
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`ture conditions and as xenografts in nude mice. lmmunocy­
`
`
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`biologic features essential to the use of these lines in a
`regularly
`�ing Cur­
`
`
`
`
`
`tochemistry using panels of antibodies raised against 6 types of
`
`
`
`disease-oriented in vitro antitumor drug screen and to the
`
`itation In­
`
`
`interpretation of data derived therefrom.
`
`
`
`intermediate filaments, 7 adenocarcinoma-associated antigens,
`Chemical
`
`
`7 melanoma/neuro-ectodermal-associated antigens, 3 neuroen­
`'Sheffield
`,ersity
`of
`
`
`
`
`docrine-associated aniigens, 9 urinary tract associated anti­
`The National Cancer Institute is implementing a new investi­
`
`
`
`:ambridge
`
`
`
`gens, and 4 markers of muscle differentiation was done on cells
`
`
`
`
`gational primary in vitro antitumor drug screen in which all
`ernational
`Reference
`
`
`
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`new agents are tested initially against a broad panel of human
`
`
`
`
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`grown in mono/ayer culture. Central nervous system (CNS)
`"NRS.
`
`
`
`
`
`
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`cell lines lacked expression of glial fibrillary acidic protein, but
`
`
`
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`tumor cell lines, arranged in a series of subpanels represent­
`nternal or
`
`
`
`all hq_d other features consistent with derivation from glioblas­
`
`
`
`ing various major categories of human cancer. An important
`1al use of
`NCER RE·
`
`
`
`
`design goal of the screen is to facilitate the detection and
`
`
`
`toma._ Lines derived from adenocarcinomas of the colon, lung
`$2.00 per
`
`and ovary, for the most part, expressed adenocarcinoma­
`
`
`
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`selection of line-specific or subpanel-specific antitumor leads
`1 to Copy·
`
`
`
`for follow-up in vivo evaluation in xenograft models em­
`
`
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`associated antigens and showed histological and/or ultra­
`,et, Salem,
`. that have
`
`
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`structural evidence of gland formation and other adenomatous
`
`
`
`
`ploying the sensitive lines identified in the primary screen.
`·c a sepa­
`
`
`
`
`The rationale, evolution and current status of this ex­
`:d: The fee
`orting Ser·
`
`
`
`
`
`
`perimental screening approach are described elsewhere (1-3).
`
`
`
`An element critical to the design of this «disease-oriented»
`
`
`
`
`
`Correspondence to: Sherman F. Stinson, Ph.D., National Insti­
`ANTICAN·
`
`
`(2)screening model is the composition of the cell line panels.
`ty for the
`
`
`tute, Building 1052, Room 121, Frederick, MD 21702-1201,
`'·
`
`
`Tumor types from �hich the lines were derived should be
`or for the
`U.S.A.
`,g herein.
`
`
`representative of well-defined human tumor classifications,
`
`
`
`and individual cell lines must faithfully reflect appropriate
`• GREECE
`
`
`Key Words: Human tumor cell lines, characterization, im­
`
`
`
`
`biological properties of the tumor of origin. To determine if
`
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`munocytochemistry, electron microscopy.
`
`
`
`0250-7005/92 $2.00+.40
`
`1035
`
`

`

`ANTICANCER RESEARCH 12: 1035-1054 (1992)
`
`these minimal criteria were met among lines initially available
`to us, and to aid in the selection of the most appropriate lines
`for interim use in a pilot screening panel, in-depth character(cid:173)
`ization of candidate lines was conducted to define various
`biologic and pharmacologic traits, and to determine their
`suitability for use under assay conditions ( 4-5).
`As a part of the above essential cell line characterization,
`detailed morphological and immunocytochemical evaluations
`also were performed. There were two major objectives of
`these evaluations. The first was to define the baseline charac(cid:173)
`teristics of the individual cell lines, to confirm their tissues of
`origin, tumor type and degree of differentiation, and to relate
`certain biological properties to those attributed to tumors of
`similar histologic origin. These are described in the present
`manuscript. The second was to define features of the cell lines
`of importance in interpreting drug responses. Some of these
`features being analyzed include multidrug resistance as well
`as other
`factors
`related
`to drug
`resistance
`(e.g.,
`topoisomerases, protein kinase C, tubulin, glutathione-S(cid:173)
`transferase, etc.), growth factor receptor expression, and
`oncogene expression. These studies are in progress and will
`be reported in a later publication. The use of the current
`panel of 60 lines in a pilot-scale antitumor screen is described
`in detail in a separate paper (5). The panel, characterized in
`detail herein, was adopted by NCI for routine screening
`operations starting in 1990 (2-3).
`
`Materials and Methods
`
`Ce/I lines and cell line cultivation. The sources of the majority of the cell
`lines used in this study have been previously reported (4). Sources of the
`remaining lines arc given in Table I. Cultivation of the lines, cryop(cid:173)
`rcscrvation and quality assurance procedures are described in detail
`elsewhere ( 4,5). All cell lines were adapted to RPM! 1640 cultufe medium
`supplemented with 5% fetai" bovine serum (heat-inactivated) and 2mM
`L-glutamine. The cultivation of human tumor cell lines under standar(cid:173)
`dized culture conditions permits comparison of morphological and im(cid:173)
`munocytochcmical features, as well as growth and drug sensitivities
`among tumor cell types.
`When possible, cell lines were also established and grown as xenog(cid:173)
`rafts in nude mice, by conventional techniqLes (15). Xenografts were
`derived by implanation of freshly thawed cell stocks, or in some cases, by
`injection of cells harvested from logarithmic growth phase cultures into
`athymic NCr-nu mice (Animal Production Area, National Cancer Insti(cid:173)
`tute, FCRDC, Frederick, MD or Taconic, Inc, Germantown, NY).
`Xcnografts of cell lines were grown subcutaneously, with the exception of
`OVCAR-3 which was cultivated intraperitoneally.
`
`'
`
`fmmunocytochemical assays of solid tumor cell lines. The anti-human
`antibodies used to characterized solid tumor cell lines, together with their
`reported antigenic specificities, working dilutions, and sources, are
`summarized in Table II. Optimal dilutions of antibodies were determined
`by titration assay against known positive and negative control samples.
`, When flask cultures were 70-90% confluent, cells were harvested with
`.a cell scraper (Costar, Cambridge, MA). Cytology specimens were
`prepared on glass slides using a Cytospin 2 cytoccntrifuge (Shandon,
`Pittsburg, PA) operated at 750 rpm for three minutes, or by application to
`Teflon coated slides containing multiple uncoated assay wells (Ccl-line
`Assoc., Inc. , Newfield, NJ). Immunostaining was performed on air-dried,
`unfixed specimens (for cell surface antigens) or on specimens fixed in
`acetone at -70°C for 1 minute (for intracellular antigens), with commer-
`
`1036
`
`Table I. Sources oj selected screening panel cell lines.
`
`Hist-0logy
`cell line
`
`CNS
`
`Reference Contributor ( Original source)
`
`SF-268
`SNB-78
`XF-498L
`
`6
`7
`8
`
`Colon adenoca.
`
`HCT-15
`
`KM-12
`
`KM-20L2
`
`9
`
`10
`
`10
`
`Lung adenoca.
`
`HOP-18
`
`HOP-62
`
`Lung-Lg. cell ca.
`
`HOP-92
`
`LXFL529L
`
`Melanoma
`
`M-14
`
`M19-MEL
`
`UACC-62
`UACC-257
`
`11
`
`11
`
`Renal carcinoma
`
`12
`
`8
`8
`13
`
`14
`
`ACHN
`
`RXF-393L
`RXF-631L
`TK-10
`
`786-0
`
`Lymphoma
`
`P.L. Kornblith
`M.L. Rosenblum, Univ. of California
`NCI/DCT/DTP•
`
`ATCC (D. L. Dexter, Roger Williams General
`Hospital)
`I.J. Fidler, Univ. of Texas M.D. Anderson
`Hospital and Tumor Institute
`I.J. Fidler , Univ. of Texas M.D. Anderson
`Hospital and Tumor Institute
`
`M. Liu, Johns Hopkins Univ. School of
`Medicine
`M. Liu, Johns Hopkins Univ. School of
`Medicine
`
`M. Liu, Johns Hopkins Univ. School of
`Medicine
`H.-H. Fiebig, Univ. ofFreiburg, West
`Germany
`
`D.H. Kern, John Wayne Cancer Clinic, UCLA
`School of Meidicine
`D.H. Kern, John Wayne CancerClinie, UCLA
`School of Meidicine
`A. Leibovitz, University of Arizona
`A . Leibovitz, University of Arizona
`
`S.M. Schmid , Southern Research Inst. (T.F.
`Hogan, Middleton Memorial VA Hospital
`NCI/OCT/DTP•
`NCI/OCT/DTP"
`R .V. Clayman, Washington Univ. School of
`Medicine
`R.D. Williams, University of Iowa
`
`..
`(
`
`SR
`
`W.J. Urba, Program Resources, Inc.
`
`"Developed under contract by Program Resources, Inc., from human,
`tumorxenografts contributed by H .-H. Fiebig, University ofFreiburg,
`West Germany
`
`~
`I I
`
`J l l I )
`
`I
`
`Table
`
`Cl
`Desig
`
`lnten
`
`AE-1
`AE-3
`Vime
`GFA
`Neun
`Desm
`
`Aden
`
`CEA
`8H12
`18D1
`47D1
`872.c
`OCl~
`19-9
`
`Mela1
`
`S-100
`R24
`85.2
`BOU
`Ll0l
`Ta99
`
`Neur,
`
`MOC
`123(
`NSE1
`
`Urim
`
`1143
`S4
`S22
`S27
`F23
`F32
`Oms
`T16
`T43
`
`Muse
`
`DE-l
`MG(cid:173)
`MY-:
`84
`
`0Abb
`recer
`gTra1
`
`

`

`--:----
`
`-
`-
`
`ieneral
`
`son
`
`son
`
`:,UCLA
`
`;, UCLA
`
`.(T.F.
`pital
`
`100l of
`
`,man
`iburg,
`
`Stinson et al: Characterization of Human Tumor Cell Lines
`
`Table II. Antibodies used for solid tumor cell line characterization.
`
`Clone/
`
`Immunizing agent/
`Specificity
`
`lg
`Subclass
`
`Derivation•
`
`Working
`dilution
`
`Source
`
`Intermediate filaments
`
`IgG1
`lgG 1
`lgG1
`lgG1
`IgG1
`IgG1
`
`lg
`lgG1k
`lgM1
`lgGlk
`IgG1
`lgG1
`IgG1
`
`IgG
`lgG3
`lgG2•
`lgG1
`lgG2a
`lgG2a
`
`lgG1
`lgG1
`lg
`
`lgG1
`lgG20
`lgG 1
`IgG1
`lgG2.
`IgM
`lgG1
`IgG2b
`lgG 1
`
`lgG1
`IgG1
`IgG1
`IgG1
`
`Mouse-asc
`Mouse-asc
`Mouse-sup
`Mouse-sup
`Mouse-sup
`Mouse-sup
`
`Rabbit-ser.
`Mouse-asc.
`Mouse-asc.
`Mouse-asc.
`Mouse-asc.
`Mouse-asc.
`Mouse-asc.
`
`Rabbi t-ser.
`Mouse-asc.
`Mouse-asc.
`Mouse-asc.
`Mouse-asc.
`Mouse-asc.
`
`1:10
`1:10
`1:10
`1:10
`1:10
`1:10
`
`1:50
`1:50
`1:50
`1:50
`1:40
`1:200
`
`1:10
`1:10
`1:10
`1:10
`1:10
`
`CRL (Cambridge, MA)
`CRL (Cambridge, MA)
`CRL (Cambridge, MA)
`CRL (Cambridge, MA)
`CRL (Cambridge, MA)
`CRL (Cambridge, MA)
`
`DuPont (Boston, MA)
`DuPont (Boston, MA)
`DuPont(Boston, MA)
`DuPont (Boston, MA)
`Centocor (Malvern, PA)
`Centocor(Malvern, PA)
`
`\
`
`Signet (Dedham, MA)
`Signet (Dedham, MA)
`Signet (Dedham, MA)
`Signet (Dedham , MA)
`Signet (Dedham, MA)
`Signet (Dedham, MA)
`
`Mouse-sup.
`Mouse-asc.
`Rabbit-ser.
`
`1:10
`1:20
`1:150
`
`Sanbio bv (Uden, Neth)
`Sanbio bv (Uden, Neth)
`ICN (Lisle, IL)
`
`Mouse-asc.
`Mouse-asc.
`Mouse-asc.
`Mouse-asc.
`Mouse-asc.
`Mouse-asc.
`Mouse-asc.
`Mouse-asc.
`Mouse-asc.
`
`Mouse-asc.
`Mouse-asc.
`Mouse-asc.
`Mouse-asc.
`
`1:100
`1:100
`1:100
`1:100
`1:100
`1:10
`1:10
`1:100
`1:10
`
`1:200
`1:400
`1:600
`1:500
`
`CRL(Cambridge, MA)
`CRL (Cambridge, MA)
`CRL(Cambridge, MA)
`CRL (Cambridge, MA)
`CRL(Cambridge , MA)
`CRL(Cambridge, MA)
`CRL (Cambridge, MA)
`CRL (Cambridge, MA)
`CRL (Cambridge, MA)
`
`ICN (Lisle, IL)
`ICN (Lisle,.IL)
`ICN (Lisle, IL)
`ICN (Lisle, IL)
`
`l
`I Designation
`• I
`I
`I
`
`I
`
`I
`
`AE-1
`AE-3
`Vimentin
`GFAPb
`Neurofila_ment
`Desmin
`
`Type I acidic keratins
`Type II basic keratins
`Vi men tin 57 Kd
`GFAP56Kd
`Neurofilament 70 & 200 Kd
`Desmin56Kd
`
`Adenocarcinoma associated surface antigens
`
`CEA
`8Hl2
`18D12
`47010
`B72.3
`OC125
`19-9
`
`Carcinoembryonic Antigen
`Cell line MCF7
`CelllineMCF7
`, CelllineA-549(GP67-98)
`Mammary Carcinoma (T AG-72)
`Cell line OVCAR-3
`Cell line SW1116
`
`Melanoma associated antigens
`
`1
`
`S-100
`R24
`B5 .2
`BOIS
`LIO!
`Ta99
`
`S-100 protein
`Cell line SK-MEL-28
`Cell line SK-MEL-93
`Cell line SK-MEL-37
`Cell line SK-MEL-33 '
`Cell line SK-MEL-23
`
`Neuroendocrine associated antigens
`
`MOC!
`123C3
`NSE1
`
`SCLC0 cells
`SCLC membrane prep.
`Bovine brain gig homodimer (90 kd)
`
`Urinary tract associated antigens
`
`J143
`S4
`S22
`S27
`F23
`F32
`Om5
`T16
`T43
`
`Cell line 253-J
`Cell line SK-RC-7
`Cell line SK-RC-7
`CelllineSK-RC-7
`Kidney epithelium (GP 140)
`CelllineSK-RC-1
`BladderTCC8
`Cell line T-43
`Cell line T-24
`
`Muscle associated antigens
`
`DE-U-10
`MG-1
`MY-32
`B4
`
`Desmin (porcine stomach)
`Human skeletal myoglobin
`Skeletal muscle myosin
`Actin ( chicken gizzard)
`
`"Abbreviations: asc. = ascites fluid; sup. = hybridomasupernatant ; ser. = whole serum bGlial fibrillary acidic protein; <Epidermal growth factor
`receptor; dParentheses enclose antigen characterization infordiation, e.g. glycoprotein, 170 kd; •small cell lung cancer; 1Neuron specificenolase;
`8Transitional cell carcinoma . .
`
`1037
`
`

`

`ANTICANCER RESEARCH 12: 1035-1054 (1992)
`
`Table Ill. Expression of intermediate filaments.
`
`Panel
`CNS
`
`Colon
`
`Lung-Ad Ca
`
`Lung-Lg cell
`
`Lung-Squamous
`Lung-Sm . cell
`
`Melanoma
`
`Ovary
`
`Kidney
`
`'
`
`Cell line
`SF-268
`SF-295
`SF-539
`SNB-19
`SNB-75
`SNB-78
`U-251
`XF-498
`COLO-205
`DLD-1
`HCC-2998
`HCT-15
`HCT-116
`HT-29
`KM-12
`KM-20L2
`SW-620
`A549
`EKVX
`HOP-18
`HOP-62
`NCI-H23
`NCI-H322
`NCI-H522
`HOP-92
`LXFL529L
`NCI-H460
`NCI-H226
`OMS 114
`OMS 273
`LOX-lMVI
`MALME-3M
`Ml4
`Ml9-MEL
`SK-MEL-2
`SK-MEL-5
`SK-MEL-28
`UACC-62
`UACC-257
`JGROV-1
`OVCAR-3
`OVCAR-4
`OVCAR-5
`OVCAR-8
`SK-OV-3
`ACHN
`A498
`CAKI-1
`RXF-393L
`RXF-631L
`SN12-C
`TK-10
`UO-31
`786-0
`
`AE-1
`0
`0
`0
`0
`0
`0
`0
`0
`50++
`95++
`100++
`100++
`95++
`100++
`100+
`100·++
`75++
`50+
`IO++
`100++
`100++
`1++
`40+
`I++
`50+
`0
`20+
`60+
`0
`· o
`0
`0
`0
`0
`0
`0
`0
`0
`0
`40+
`90+
`80+
`80+
`0
`40+
`95 ++
`75+
`25+
`0
`0
`75++
`95++
`90++
`50++
`
`AE-3
`0
`0
`0
`0
`0
`0
`0
`0
`50++
`100++
`0
`95++
`100++
`50++
`100++
`100+
`50++
`100++
`75++
`100++
`100++
`1++
`30+
`0
`0
`0
`40++
`100++
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`60+
`70+
`90+
`100+
`0
`15+
`50 ++
`0
`0
`0
`0
`100++
`0
`0
`0
`
`Vimentin
`100++
`50+
`75++
`100++
`100++
`100+ +
`100++
`100++
`0
`0
`0
`0
`0
`0
`0
`0
`75++
`100+
`75++
`25++
`90++
`100+ +
`0
`75++
`100++
`100+
`100+ +
`100+
`70+
`90+
`100+
`100++
`100++
`100++
`100++
`100+++
`100++
`100++
`100+ +
`70++
`0
`o·
`0
`100+
`100+
`100+++
`100+ +
`100++
`100++
`100+++
`100++
`95++
`100++
`80+
`
`GFAP
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`100
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`
`Neurofilament
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`n
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`
`Desmin
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`100++
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`
`cially available kits using the peroxidase anti-peroxidase technique (Sig-
`net Laboratories, Inc., D edham, MA) the avidin-biotin complex method
`(Vector Labora tories, Inc. , Burlingame, CA), or alkaline phosphatase
`methods (BIO/CAN Scientific, Portland , ME). The three techniques
`gave comparab le results. Negative controls co nsisting of isolypic im-
`munoglobulin or ascitic fluid at a similar protein concentration to the
`
`primary rmtibody were run for each smnplc . '!ides were prepared from 11
`nlinirnum of 2 cparatc cultures for each ecll linc . Microscopicall}',,lhrcc
`separate fie lds, each consL ting of approximmcly 100 cells. were selected
`randomly from c.ich slide , und the percentage of po itivcly. mince.I ;ells
`was cou nted. In 11ddi1io n, the relative , l:lli ning intensity was qunlin11ivcly
`assessed on a .calc of + 10 +:t-+, with + rcprcscnling,dctcctablc swinin&·
`
`1038
`
`I
`
`)
`
`Table
`
`Panel
`
`CNS
`
`Colon
`
`Lung-,
`
`Lung-[
`
`Lung-S
`Lung-S
`
`Mclane
`
`Ovary
`
`Kidney
`
`-
`~ I and++
`\
`I
`i frorn an
`I
`I
`
`al! field:
`nearest
`
`Histo/og
`formed ,
`
`

`

`Stinson et al: Characterization of Hum an Tumor Cell Lines
`
`Table IV. Expression of adenocarcinoma associated antigents.
`
`(
`
`Panel
`
`I CNS
`'
`I
`r
`
`I
`
`\
`
`Cell line
`SF-268
`SF-295
`SF-539
`SNB-19
`SNB-75
`SNB-78
`U-251
`XF-498
`COLO-205
`DLD-1
`HCC-2998
`HCT-15
`HCT-ll6
`HT-29
`KM-12
`KM-20L2
`SW-620
`A549
`EKVX
`HOP-18
`HOP-62
`NCI-H23
`NCI-H322
`· NCI-H522
`HOP-92
`LXFL529L
`NCI-H460
`NCI-H226
`DMS 114
`DMS 273
`LOX-IMVI
`MALME-3M
`M14
`M19-MEL
`SK-MEL-2
`SK-MEL-5
`SK-MEL-28
`UACC-62
`UACC-257
`IGROV-1
`OVCAR-3
`OVCAR-4
`OVCAR-5
`OVCAR-8
`SK-OV-3
`ACHN
`A498
`CAKI-1
`RXF-393L
`RXF-631L
`SN12-C
`TK-10
`UO-31
`786-0
`
`CEA
`0
`0
`0
`0
`0
`0
`0
`0
`100++
`25++
`0
`0
`0 .
`100++
`100++
`100++
`1++
`10++
`2++
`100++
`0
`0
`0
`l++
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`1++
`100++
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`
`8H12
`0
`0
`0
`0
`0
`0
`0
`0
`100++
`70++
`25++
`0
`0
`100++
`0
`50++
`10++
`0
`0
`10++
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`20++
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`
`18D12
`0
`0
`0
`0
`0
`0
`0
`0
`100+
`80++
`95+
`100++
`0
`100++
`80++
`0
`20++
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`100++
`100+
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`
`47D10
`0
`0
`0
`0
`0
`0
`0
`0
`100+
`5++
`5+
`0
`0
`100+
`50++
`100++
`50+
`5++
`10++
`100++
`0
`0
`10++
`5++
`100+
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`100+
`0
`0
`0
`JOO++
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`
`872.3
`0
`0
`0
`0
`0
`0
`0
`0
`5+
`5+
`10+
`0
`0
`0
`20+
`5+
`0
`5+
`15+
`5+
`0
`0
`5+
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`20+
`75+
`25+
`0
`25+
`0
`0
`0
`0
`0
`0
`0
`·o
`0
`
`OC125
`0
`0
`0
`0
`0
`0
`0
`0
`10+
`0
`75++
`0
`0
`0
`0
`0
`0
`0
`95++
`35+
`0
`0
`60+
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`5+
`65++
`35+
`45+
`0
`5+
`0
`0
`· 0
`0
`0
`0
`0
`0
`0
`
`19-9
`0
`0
`0
`0
`0
`0
`0
`0
`100+++
`80+++
`35++
`0
`0
`60+ + +
`0
`45+++
`20++
`0
`0
`50+++
`0
`0
`0
`0
`0
`0
`0
`0
`0
`
`0
`5+
`0
`0
`5+
`0
`0
`0
`0
`0
`25++
`0
`95+++
`0
`10+
`0
`5+
`30+
`0
`0
`0
`0
`15+
`0
`
`Colon
`
`I
`'
`I
`(
`
`Lung-Ad Ca
`
`l
`
`Lung-Lg cell
`
`Lung-Squamous
`Lung-Sm. cell
`
`Melanoma
`
`Ovary
`
`Kidney
`
`f
`
`from a
`1, three
`. elected
`ed cells
`tativel y
`1aining,
`
`l
`
`•' I and++ strong and++ + very strong staining respectively. Results from
`\
`
`all fields and slides for each cell line were averaged and rounded to the
`nearest 5%.
`
`fli.1·10/ogy and hisruchemislry . Histology and histochemistry were per-
`formed on xe_nografts of the cell lines. Tumors (0.5-1.0 cm) were removed
`
`\
`
`11
`
`• frmn ~nimals, fixed in 10% neutral buffered formalin and cmbcddc\J in
`I
`I
`
`paraffin. Five µm thick sections were stained with hematoxylin-eosin ,
`alcian blue-PAS (for differentiation of mueopolysaecharides), Kreyberg
`stain (for keratin), Lilic stain (for melanin), and Bodian stain (for
`argyrophilic substances) .
`
`Electron microscopy. Ultrastructural examination was performed on solid
`tumor cell lines grown in monolayer culture and on xcnografts of cell
`
`1039
`
`

`

`ANTICANCER RESEARCH 12: 10~5-1054 (1992)
`
`Table V. Expression of melanoma associated antigens.
`
`Panel
`CNS
`
`Colon
`
`Lung-Ad Ca
`
`Lung-Lg cell
`
`Lung-Squamous
`Lung-Sm. cell
`
`Melanoma
`
`Ovary
`
`' Kidney
`
`Cell line
`SF-268
`SF-295
`SF-539
`SNB-19
`SNB-75
`SNB-78
`U-251
`XF-498
`COL0-205
`DLD-1
`HCC-2998
`HCT-15
`HCT-116
`HT-29
`KM-12
`KM-20L2
`SW-620
`A549
`EKVX
`HOP-18
`HOP-62
`NCI-H23
`NCI-H322
`NCI-H522
`HOP-92
`LXFL529L
`NCI-H460
`NCI-H226
`DMS 114
`DMS 273
`LOX-IMVI
`MALME-3M
`M14
`M19-MEL
`SK-MEL-2
`SK-MEL-5
`SK-MEL-28
`UACC-62
`UACC-257
`IGROV-1
`OVCAR-3
`OVCAR-4
`OVCAR-5
`OVCAR-8
`SK-OV-3
`ACHN
`A498
`CAKI-1
`RXF-393L
`RXF-631L
`SN12-C
`TK-10
`U0-31
`786-0
`
`S-100
`0
`0
`0
`0
`65++
`0
`0
`100+++
`0
`0
`0
`0
`0
`0
`0
`0
`
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`70+++
`50+
`60+
`45+
`0
`0
`0
`5+
`0
`0
`0
`0
`0
`35+
`0
`0
`0
`0
`0
`0
`0
`0
`0
`
`R24
`0
`0
`0
`0
`0
`0
`0
`70+
`0
`0
`0
`0
`0
`0
`0
`0
`
`0
`0
`40+
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`5+
`90++
`45+
`45+
`45+
`60+
`80+
`60+
`30+
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`
`B5.2
`50++
`0
`10+
`0
`65+
`80+
`0
`85++
`0
`0
`25+
`0
`0
`0
`0
`0
`
`0
`0
`30+
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`10+
`95++
`90++
`75+++
`40++
`0
`30+
`80+
`55++
`0
`35+
`0
`0
`90+
`0
`0
`0
`0
`0
`0
`50+
`0
`0
`0
`
`BD18
`0
`5+
`0
`0
`70+++
`0
`0
`75++
`0
`0
`0
`0
`0
`0
`0
`0
`
`0
`0
`10+
`0
`0
`0
`0
`25+
`0
`0
`0
`0
`0
`0
`50+
`85++
`10++
`60++
`25+
`65+
`45+
`15+++
`0
`0
`0
`25+
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`
`LlOl
`45+
`60+
`10+
`60+
`80+
`65+
`40+++
`100+++
`75+
`25+
`70+
`0
`0
`0
`50+
`0
`85+
`50+
`85++
`0
`85++
`0
`0
`0
`50++
`10+
`55++
`5+
`0
`25+
`0
`100+++
`5++
`75+++
`55+++
`75+++
`90+++
`90+
`60+++
`0
`70+
`70+
`35+
`100++
`35+
`95++
`5+
`80+++
`85++
`20+
`40+
`80+
`95+++
`95+
`
`Ta99
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`50+
`0
`35+
`0
`40+++
`10+
`0
`90+++
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`
`lines. For in vitro lines, cells were scraped from culture vessels after the
`monolaycr had reached 70-90% conOuency. The cell were edlmcntcd
`into pellets· by centrifugation O.l approximately 200 x g for 10 minutes,
`and fixed in .1,25% glu1araldehydc in 0.1 M cucodylatc buffer (pH = 7.2)
`for I hour. For .xenografts, several piece of ti sue ranging in diameter
`from 0.5 to 'I mm were cut from non-necrotic areas and lixcd for 24 hours
`in 2.5% glutaraldehyde in 0.1 M cacodylate buffer. Cell pellets and tissue
`pieces were post-fixed in osmium tetroxide, dehydrated in graded solu-
`
`lions of ethanol and embedded in Epon 812 resin (Fluka AG, Ducks,
`wit:zcrland). Uitrathin sections were 1aincd with uranyl acetate und l~ 3d
`citrate and examined with a JEOL JEM- IOO X c.lectron. microscope using
`an accelerating voltage of 60 KV.
`
`Flow cytometry of le11kemia and lymphoma cel/ li11e.s. Phcnotypic clmruc-
`rcrization of the cell liaes K-562, MOLT-4, HL-60, CCR f'-CEM, · RP~l
`8226 and SR were conducted by flow cytometry . Cells were uspen led 10
`
`Table
`Panel
`
`Colon
`
`)
`
`I
`I
`I
`
`I
`
`f
`
`Lung-
`
`Lung-
`
`Lung-.
`Lung-
`
`Melan
`
`Ovary
`
`Kidne
`
`phospl
`humar
`for C
`antibo
`CD57.
`CD14.
`antibo
`
`,I
`
`I
`
`l
`
`1040
`
`I
`I
`
`

`

`Panel
`
`Colon
`
`Lung-Ad Ca
`
`Lung-Lg cell
`
`Lung-Squamous
`Lung-Sm. cell
`
`Melanoma
`
`Ovary
`
`Kidney
`
`Table VJ. Erpre.ssim1 of neuroendocrine associated a111rge11s.
`Cell line
`NSE
`5+
`SF-268
`SF-295
`0
`0
`SF-539
`SNB-19
`0
`SNB-75
`0
`SNB-78
`30+
`U-251
`70+
`XF-498L
`0
`0
`COLO-205
`0
`OLO-1
`0
`HCC-2998
`0
`HCT-15
`0
`HCT-116
`0
`HT-29
`KM-12
`0
`0
`KM-20L2
`0
`SW-620
`0
`A549
`65+
`EKVX
`0
`HOP-18
`0
`HOP-62
`0
`NCI-H23
`0
`NCI-H322
`NCI-H522
`0
`HOP-92
`0
`LXFL529L
`0
`NCI-H460
`0
`NCJ-H226
`0
`OMS 114
`0
`OMS 273
`0
`0
`LOX-IMVI
`70+
`MALME-3M
`0
`M14
`0
`M19-MEL
`0
`SK-MEL-2
`20+
`SK-MEL-5
`SK-MEL-28
`0
`0
`UACC-62
`UACC-257
`45++
`0
`IGROV-1
`0
`OVCAR-3
`0
`OVCAR-4
`0
`OYCAR-5
`0
`OVCAR-8
`0
`SK-OV-3
`0
`ACHN
`0
`A498
`0
`CAKI-1
`0
`RXF-393L
`0
`RXF-631L
`40+
`SN12-C
`0
`TK-10
`0
`UO-31
`0
`786-0
`
`Stinson et al: Characterization of Human Tumor Cell Lines
`
`\
`
`MOC 1
`25+
`0
`0
`25+
`40+++
`35+
`80++
`50+
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`85++
`55++
`0
`0
`5+
`0
`0
`5+
`45+
`0
`0
`0
`60+
`40+
`75+++
`45++
`65++
`0
`0
`0
`0
`0
`0
`55++
`0
`0
`0
`55++
`10+
`
`123C3
`30+
`0
`0
`50+
`75+++
`50+++
`90++
`95++ +
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`95+++
`25++
`0
`0
`5+
`0
`0
`5+
`80+ ++
`5++
`10+
`0
`50++
`50++
`75++
`75++
`75++
`0
`0
`25+
`50++
`0
`O·
`75++
`0
`0
`0
`50+++
`50+++
`
`obtained from Becton-Dickinson lmmunocy lometry Systems (Mountain
`phosphate buffered saline (PBS) containing 2% heat-inactivated pooled
`View, CA). FITC-conjugated anti-CD?, CD116, CD29 and CD45RA
`human AB serum for 5 minutes to block Fe receptors or in PBS-2% BSA
`for CD16 (Fe receptor III) staining. Fluorescein (FITC)-conjugatcd were obtained from Coulter Corporation (Hialeah, FL) . FITC-conju-
`gated an tiglycophorin and unconjugated anti-CD18 were obtained from
`antibodies to CO2, CD3, CO4, CDS , CDlO, CD16, CD19, CD25, CD45 ,
`CD57 , and CD71; phycoerythrin (PE)-conjugated antibodies to CD8, GcnTrak, Inc. (Plymouth Meeting, PA). Cells incubated with unconju-
`gated primary antibodies were washed and incubated with fluorescein-
`CD14, CD23, CO33, CD38, CD56 and HLA-OR; and uneonjugated
`conjugated goat anti-mouse IgG (Tago, Inc. , Burlingame, CA). Stained
`antibodies to human IgG, lgM, kappa and lambda light chains were
`
`1041
`
`- - . ,
`
`r
`
`I
`
`\
`
`-
`
`G , Bucks ,
`le and \cad
`.cope using
`
`pie charac-
`M, RPM!
`spended in
`
`I
`
`I
`I
`
`

`

`ANTICANCER RESEARCH 12: 1035-1054 (1992)
`
`Tabl~ VII. Expression of urinary tract associated antigens.
`
`Panel
`
`CNS
`
`Colon
`
`Lung-Ad Ca
`
`Lung-Lg Cell
`
`Lung-Squamous
`Lung-Sm. Cell
`
`Melanoma
`
`Ovary
`
`Kidney
`
`Cell line
`
`1143 .
`
`S4
`
`S22
`
`S27
`
`F23
`
`F31
`
`OmS
`
`T16
`
`T43
`
`0
`70++
`SF-268
`0
`95++
`SF-295
`0
`55 ++
`SF-539
`0
`95++
`SNB-19
`30+
`85 ++ +
`SNB-75
`0
`40++
`SNB-78
`U-251
`0
`95+++
`XF-498L
`0
`95+++
`COLO-205
`0
`100+
`DLD-1
`10+
`100++
`HCC-2998
`0
`75 ++
`0
`85++
`HCT-15
`0
`HCT-116
`0
`HT-029
`95++
`0
`0
`KM-12
`0
`KM-20L2
`95+++
`5+
`SW-620
`0
`10+
`A549
`100++ + 0
`EKVX
`85++
`0
`HOP-18
`80+++
`0
`HOP-62
`90+ ++
`0
`20+
`NCI-H23
`0
`85++
`NCI-H322
`0
`0
`NCI-H522
`0
`HOP-92
`60+++
`0
`0
`LXFL529L
`0
`40++
`NCI-H460
`0
`NCI-H226
`100++ + 0
`DMS 114
`0
`0
`DMS 273
`85++
`0
`LOX-IMVI
`JOO+++ 0
`MALME-3M 90+++
`0
`Ml4
`85++
`0
`M19-MEL
`80++
`0
`SK-MEL-2
`40++
`0
`SK-MEL-5
`15+ +
`0
`SK-MEL-28
`55+ +
`0
`UACC-62
`25++
`0
`UACC-257
`55+ +
`0
`IGROV-1
`80+++
`0
`OVCAR-3
`55 ++ +
`0
`OVCA R-4
`90++
`0
`OVCAR-5
`95++
`0
`OVCAR-8
`90++
`0
`SK-OV-3
`90+++
`20+
`A498
`45 ++
`0
`ACHN
`100+++ 0
`CAKI-1
`85+++
`20+
`RXF-393L
`50+ +
`20++
`RXF-631L
`80++
`0
`SN12-C
`85+++
`0
`TK-10
`85+++
`60+
`U0-31
`85+++
`0
`786-0
`85 ++ +
`90+++
`
`0
`0
`0
`0
`60+++
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`so++
`0
`0
`0
`70++
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`s +
`95+++
`35++
`0
`0
`80++ +
`50+
`55++
`
`0
`0
`0
`0
`0
`0
`0
`0
`70+
`10+
`35++
`0
`0
`0
`85+++
`s+
`0
`0
`0
`0
`0
`0
`0
`0
`10+
`0
`0
`70+
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`70+++
`5+
`0
`0
`0
`25++
`25++
`90++
`15+
`50+ +
`5+
`0
`75++
`0
`95++
`
`0
`0
`25++
`0
`25+
`15+
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`10+
`5+
`10+
`30+
`0
`0
`0
`20+
`0
`0
`0
`0
`s+
`so++
`0
`0
`0
`0
`0
`0
`0
`0
`0
`5+
`0
`0
`55+
`80++
`IO+
`0
`70+++
`65++
`5+
`80++
`0
`45+
`85++
`
`s+
`55++
`0
`0
`85+
`0
`30+
`0
`90+
`25+
`0
`95+
`80+
`35+
`60++
`0
`70+
`95+
`0
`0
`80+
`50+
`65+
`0
`25+
`0
`40+
`100+
`0
`20+
`0
`0
`0
`65++
`40+
`75++
`60+
`0
`65+
`50+
`0
`0
`· 80+
`100+
`60++
`95++
`10+
`85+
`35+
`0
`0
`0
`80+
`95 + +
`
`0
`so++
`0
`0
`65+
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`5+
`0
`5+
`80+
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`60++
`0
`0
`0
`0
`0
`45++
`0
`0
`0
`0
`0
`0
`0
`0
`5+
`0
`0
`
`0
`0
`0
`0
`0
`0
`0
`0
`50+
`80++
`0
`45++
`0
`95++
`0
`65 + +
`0
`0
`0
`0
`0
`0
`85+++
`0
`32++
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`25 ++
`85++
`95++
`0
`80++
`0
`0
`15+
`0
`0
`0
`0
`0
`0
`
`55++
`70+++
`20++
`70+++
`85+++
`90++
`50+++
`75+
`100+++
`70+++
`75+++
`75+++
`90+++
`80+++
`90+++
`40+++
`100+++
`100+++
`95+++
`80++
`85+++
`75++
`95+++
`40+
`55++
`60+
`95 ++
`95++
`45+
`25++
`90+
`95++
`95++
`70+++
`70++
`65+++
`85+++
`95++
`70+++
`55++
`75+++
`75+++
`90+++
`95 ++
`60++
`100+
`50+++
`75+++
`80+++
`95+++
`55+
`95++
`85++
`100+++
`
`cells were washed in protein-free phosphate buffered saline, fixed with
`buffered I% paraformaldehyde and stored in the dark at 4°C overnight
`prior to analys is. Flow cytomctric analysis of cells was performed on a
`Coulter Profile flow cytometer (Hialeah, FL) with 4 decade log amplifica-
`tion.
`
`Viable cell. ·were bitmap guted bn cd on light ·cnttcr (Qr nnalysi . . Two
`color immunolluore cent
`·mnplcs were co1Tcetcd for spectral overlap
`using color compensation . The photomultiplier tube (PMT) vollogc u cd
`t nnalyzc fluorescence si lt1ols was adjusted for cilch ccll lin bas~ on the
`u1.>ch1
`intensity of autofluorcs nee obtained on nuorcsccntly labelled
`
`1042
`
`I
`
`I
`I
`
`Table
`
`Cell I
`
`CNS
`SF-26
`
`SF-29
`
`SF-53
`
`SNB-
`
`SNB-
`
`SNb-
`
`U-25
`
`XF-4
`
`COL
`COL
`
`DLD
`
`HHC
`
`HCI
`
`HCI
`
`HT<
`
`KM-
`
`KM-
`
`SW-•
`
`Lm
`A54'
`
`EK,
`
`HOI
`
`HOI
`
`NCI
`
`NCI
`
`vsM
`
`LUI
`HO
`
`LXI
`
`NCI
`
`

`

`-
`-
`
`+
`
`+
`+
`
`Stinson et al: Characterization of Human Tumor Cell Lines
`
`Table VIII. Selected ultrastructura/ features of cell lin~s cultivated in mono/ayer culture and as xenografls in nude mice.
`
`CellrTissue
`organizations
`
`\
`
`Cell line
`
`CNS
`SF-268
`
`SF-295
`
`SF-539
`
`SNB-19
`
`SNB-75
`
`SNb-78
`
`U-251
`
`XF-498
`
`COLON
`COLO-205
`
`DLD-1
`
`HHC-2998
`
`HCT-15
`
`HCT-116
`
`HT-29
`
`KM-12
`
`KM-20L2
`
`Cultivation
`method
`
`RER'
`
`Golgi
`
`Inclusions
`
`Surface
`features
`
`in vitro
`in vivod
`in vitro
`in vivo
`in vitro
`in vivod
`in vitro
`in vivo
`in vitro
`in vivod
`in vitro
`in vivod
`in vitro
`in vivo
`in vitro
`in vivo
`
`+b
`
`+
`+
`+
`
`+
`+++
`++
`
`++
`
`+
`+++
`++
`++
`
`++fpC
`
`+ ++fp
`
`++fp
`
`+++fp
`+fp
`+++fp
`
`++fp
`
`++fp
`
`+++fp
`++fp
`
`mucous
`
`mucous
`
`mucous
`mucous
`
`mucous
`mucous
`
`+
`
`+
`
`+
`+
`+
`
`+
`
`+
`++
`+
`+
`+
`
`+dis
`
`+dis
`
`+dis
`
`+dis
`
`+dis
`+dis
`+dis
`
`+++mv
`
`+mv
`+mv
`++mv
`
`+mv
`
`++fp
`++mv
`++mv
`++mv
`++fp
`
`Junctions
`
`groups
`
`groups
`
`groups
`
`groups
`
`chords
`chords
`chords
`
`+ ds,za
`++ ds
`++ds
`++ds
`++ds
`++ds
`++ds
`+ds
`++ds
`+++ds
`++ds,za
`++ds,za
`+ds
`
`in vitro
`in vivo
`in vitro
`in vivo
`in vitro
`in vivo
`in vitro
`in vivo
`in vitro
`in vivo
`in vitro
`in vivo
`in vitro
`in vivo
`in vitro
`in vivo
`in vitro
`in vivo
`
`in vitro
`in vivo
`in vitro
`in vivo
`in vitro
`in vivod
`in vitro
`in vivod
`in vitro
`in vivod
`in vitro
`in vivo
`
`in vitro
`in vivo
`
`in vitro
`in vivo
`in vitro
`in vivo
`in vitro
`in vivo
`
`+
`+
`+
`+
`++
`+
`+
`+
`+
`++
`+
`+
`+
`+
`+
`+
`+
`++
`
`+
`+++
`+
`+
`+
`
`+
`
`+
`
`+
`++
`
`+
`++
`
`++
`+
`+
`+
`
`+
`
`SW-620
`
`LUNG-AD CA
`A549
`
`EKVX
`
`HOP-18
`
`HOP-62
`
`NCI-H23
`
`NCI-H322
`
`vsMCI-H522
`
`LUNG-LG CELL
`HOP-92
`
`LXFL529L
`
`NCI-H460
`
`r
`
`r
`
`r
`r
`r
`
`t-
`t-
`t-
`t-
`
`+
`
`-
`
`lysis. Two
`,I overlap
`ltage used
`scs on the
`:I subclass
`
`+
`
`++
`
`+
`+
`++
`
`++
`
`+
`
`mucous
`
`mucous
`
`mucous
`mucous
`
`mucous
`
`mucous
`
`+
`+
`++
`++
`+++
`+
`
`mf
`
`++mv
`+mv
`+fp
`+mv
`
`+fp
`++mv
`++mv
`+mv
`++fp
`
`++fp
`
`++mv
`
`++mv
`+mv
`
`+fp
`+mv
`
`+fp
`
`+fp,mv
`+fp,mv
`++mv
`+mv
`
`++ds,za
`+ds,za
`+ds
`+ds
`
`+ds
`++ds
`++ ds,za
`++ds
`++ds
`
`++ds
`
`+tj
`
`+ds,za
`+++ds
`
`++ds
`++ds
`
`+dis
`+tj
`
`sheets/glands
`sheets/glands
`sheets
`sheets
`chords/sheets
`chords/sheets
`sheets
`sheets
`chords/sheets/glands
`chords/sheets/glands
`chords/she