In vitro confirmation of newly established lung cancer cell lines using flow cytometry and multicellular tumor spheroids

被引:8
作者
Inoue, S
Takaoka, K
Endo, T
Mizuno, S
Ogawa, Y
Yoshida, M
Ohnuma, T
机构
[1] NIKKO MEM HOSP, DEPT RESP DIS, BOKAI, MURORNA, JAPAN
[2] HOKKAIDO UNIV HOSP, DEPT PATHOL, KITA KU, SAPPORO, HOKKAIDO 060, JAPAN
[3] HOKKAIDO UNIV, DENT HOSP, SCH DENT, CENT CLIN LAB, KITA KU, SAPPORO, HOKKAIDO 060, JAPAN
[4] HOKKAIDO UNIV, SCH MED,CENT RES INST,DIV ELECTRON MICROSCOPY, KITA KU, SAPPORO, HOKKAIDO 060, JAPAN
[5] HOKKAIDO UNIV, GRAD SCH ENVIRONM EARTH SCI, DEPT CYTOGENET, KITA KU, SAPPORO, HOKKAIDO 060, JAPAN
[6] MT SINAI SCH MED, DEPT MED NEOPLAST DIS, NEW YORK, NY USA
关键词
cytopathological type; tumor associated antigen; DNA; karyotype; electron-micrograph; flow cytometry; multicellular tumor spheroids;
D O I
10.1016/S0169-5002(97)00652-1
中图分类号
R73 [肿瘤学];
学科分类号
100214 ;
摘要
We report on a simplified method of cytomorphological in vitro confirmation of newly established lung cancer cell lines by using multicellular tumor spheroids (MTS) and flow cytometry (FCM). Eleven cell lines were established from 11 patients with lung cancer. The MTS were produced by culturing cells in agar-coated dish. Cytomorphological studies were made using smears of crushed MTS and frozen sections of MTS. The MTS were fixed doubly with paraformaldehyde and osmic acid for scanning and transmission electron microscopy. Bivariate fluorescence: of fluorescein isothyocyanate (FITC, tumor associated antigen, TAA) and propidium iodide (DNA) were measured by FCM. The MTS grew anchorage-independently. Cytopathological and electron microscopic findings of MTS were similar to those of the original clinical specimens. The DNA index and TAA were useful in evaluating the presence or absence of contamination by cells of non-tumor origin. The new cell lines satisfied a minimum of four conditions to confirm their establishment: (a) they originated from humans, (b) they were cytomorphologically identified with specimens from primary lesions, (c) they showed tumorigenicity, and (d) they were free from contamination by cells of different origin. From these findings, the establishment of new cell lines can be confirmed in vitro by using MTS and FCM. (C) 1997 Elsevier Science Ireland Ltd.
引用
收藏
页码:85 / 101
页数:17
相关论文
共 36 条
[1]  
BERGH J, 1985, AM J CLIN PATHOL, V84, P1
[2]   EXPRESSION OF C-ERBB-2 PROTEIN, NEURON-SPECIFIC ENOLASE AND DNA FLOW-CYTOMETRY IN LOCALLY ADVANCED TRANSITIONAL CELL-CARCINOMA OF THE URINARY-BLADDER [J].
BERNER, A ;
JACOBSEN, AB ;
FOSSA, SD ;
NESLAND, JM .
HISTOPATHOLOGY, 1993, 22 (04) :327-333
[3]  
CARNEY DN, 1984, CANCER RES, V44, P5399
[4]  
CARNEY DN, 1983, CANCER RES, V43, P2806
[5]   VARIABLE RADIOBIOLOGICAL RESPONSES OF SPHEROIDS [J].
DURAND, RE .
RADIATION RESEARCH, 1980, 81 (01) :85-99
[6]   CELL-CYCLE KINETICS IN AN INVITRO TUMOR MODEL [J].
DURAND, RE .
CELL AND TISSUE KINETICS, 1976, 9 (05) :403-412
[7]   RADIATION SURVIVAL OF CELLS FROM SPHEROIDS GROWN IN DIFFERENT OXYGEN CONCENTRATIONS [J].
FRANKO, AJ ;
SUTHERLAND, RM .
RADIATION RESEARCH, 1979, 79 (03) :454-467
[8]  
FRESHNEY RI, 1987, CULTURE ANIMAL CELLS, P169
[9]   HETEROTRANSPLANTATION OF HUMAN BREAST CARCINOMAS IN NUDE-MICE - CORRELATION BETWEEN SUCCESSFUL HETEROTRANSPLANTS, POOR PROGNOSIS AND AMPLIFICATION OF THE HER-2 NEU ONCOGENE [J].
GIOVANELLA, BC ;
VARDEMAN, DM ;
WILLIAMS, LJ ;
TAYLOR, DJ ;
DEIPOLYI, PD ;
GREEFF, PJ ;
STEHLIN, JS ;
ULLRICH, A ;
CAILLEAU, R ;
SLAMON, DJ ;
GARY, HE .
INTERNATIONAL JOURNAL OF CANCER, 1991, 47 (01) :66-71
[10]  
*ICSN, 1985, CELL GENET