agar and Urinary-Bladder-Neoplasms

In vitro chemotherapy sensitivity testing for transitional cell carcinoma and other urologic malignancies is an exciting research tool of great future promise. It is not yet an established clinical test of proved value for patients with genitourinary malignancies.

Topics: Agar; Carcinoma, Transitional Cell; Cell Division; Cells, Cultured; Colony-Forming Units Assay; Humans; Tumor Stem Cell Assay; Urinary Bladder Neoplasms

The aim of this study was to investigate the effect of retinoic acid (RA) and its complexes with transition metals on the bladder cancer cell line EJ. Retinoic acid complexes with transition metals Cu, Co, Zn, and Ni were prepared. Cell proliferation was tested by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay in the presence of RA or its complexes with transition metals Cu, Co, Zn, and Ni ¿Cu(RA)2.3H2O, Co(RA)2.3H2O, Zn(RA)2.4H2O, and Ni(RA)2.3H2O¿. Colony formation in soft agar culture, A agglutination reaction, and lactic acid dehydrogenase isoenzyme assay were performed in the cells treated with these drugs to estimate the induced differentiation. p53 or c-Ha-ras expression in drug-treated cells was assayed by ABC immunocytochemistry technique. The results demonstrate that EJ cells treated with the drugs become less confluent and tend to exhibit normal characteristics. Although RA and its complexes showed inhibition to proliferation of EJ cells at the concentrations of 10(-6) mmol/l, the inhibition induced by Ni(RA)2.3H2O was much more marked than that by RA. EJ cells were growth inhibited by RA or Ni(RA)2.3H2O from 48 to 96 h at the concentration of 10(-8) mol/l. The levels of LDH4 and LDH5 in the cells were greatly increased by RA. Nevertheless, Ni(RA)2.3H2O did not affect LDH isoenzyme in EJ cells. The number of colony formations of EJ cells in soft agar culture was decreased by RA or Ni(RA)2.3H2O. The percentage of colony formation in soft agar culture was much lower in EJ cells treated with Ni(RA)2.3H2O than with RA. The required concentration of A agglutination reaction was more increased for EJ cells treated with RA or Ni(RA)2.3H2O than for the control and was further increased in cells treated with Ni(RA)2.3H2O. Mutant p53 expression was more decreased in the EJ cells treated with RA or Ni(RA)2.3H2O than in the control. Although RA at the concentration of 10(-6) mmol/l caused lower p21 expression, Ni(RA)2.3H2O did not affect p21 expression in EJ cells. Therefore, RA and its transition metal complexes have a potential use in the treatment of bladder cancer.

Topics: Agar; Agglutination Tests; Antineoplastic Agents; Carcinoma, Transitional Cell; Cell Differentiation; Cell Division; Cobalt; Copper; Gene Expression Regulation, Neoplastic; Humans; In Vitro Techniques; Isoenzymes; L-Lactate Dehydrogenase; Metals; Mutation; Nickel; Proto-Oncogene Proteins p21(ras); Tretinoin; Tumor Cells, Cultured; Tumor Suppressor Protein p53; Urinary Bladder Neoplasms; Zinc

To evaluate the predictive value for recurrences of repeated urine cell culture in double layer soft-agar in patients with transitional cell carcinoma, 272 urine samples were collected from 75 patients. After a limited period of follow-up (mean 19.5 +/- 15.1 months) nine out of 46 patients with at least one evaluable culture and evaluable for clinical follow-up had a histologically proven recurrence preceded or accompanied by tumor colony growth in culture; one patient had tumor recurrence with growth negative urine cell culture; 15 patients had growth negative urine cell cultures and at first evaluation no recurrence of transitional cell carcinoma; 21 patients had growth positive urine cell cultures and no recurrence at first evaluation of follow-up. These two latter groups were evaluated again after an additional 30 months of follow-up. In the group with growth positive urine cell cultures (21 patients) nine patients (47.6%) developed a recurrence; 10 developed no recurrence. In the other group of patients with growth negative urine cell cultures (15 patients) six patients (40%) developed a recurrence, three following a growth positive urine cell culture. In the group of 46 evaluable patients in this study, 25 patients developed a recurrence, 21 had at least one growth positive culture. The sensitivity is 84% (21/25). Of 33 patients with positive cultures 21 have had a recurrence (64%) while 12/33 (36.7%) are "false positive." The false negative rate is 36.4% (four of 11 patients developed recurrences not preceded by growth positive urine cell cultures).(ABSTRACT TRUNCATED AT 250 WORDS)

Topics: Agar; Carcinoma, Transitional Cell; Cells, Cultured; Follow-Up Studies; Humans; Neoplasm Recurrence, Local; Risk Factors; Time Factors; Tumor Stem Cell Assay; Urinary Bladder Neoplasms

The in vitro clonogenicity of 25 human tumours was compared in two simple two layer culture systems, agar/agar and liquid medium/agar. There was a strong correlation between the values for clonogenicity obtained in each system. A linear relationship between cells plated and colonies formed was found in both systems. Radiation survival in the liquid culture system was essentially log linear with a small initial shoulder confirming that we were not simply counting clumps. We present a simple method of assessing the self-renewal capability of the clonogenic cells of human solid tumours, based on the liquid/agar two-layer system, which we have used to compare secondary plating efficiency and colony size analysis as measures of self renewal in human transitional cell carcinoma of the bladder.

Topics: Agar; Carcinoma, Transitional Cell; Cell Count; Cell Survival; Clone Cells; Colony-Forming Units Assay; Culture Media; Dose-Response Relationship, Radiation; Humans; Neoplasms; Tumor Stem Cell Assay; Urinary Bladder Neoplasms

Because in vitro cell growth of transitional cell carcinoma explants and cell lines often fail to adequately proliferate in semisolid media, we have examined the effect of agents used to make media semisolid (methyl cellulose, Bacto-agar, Sea Plaque agarose and Sea Prep 15/45 agarose) on the in vitro growth of 11 transitional cell carcinoma cell lines. The growth of human transitional cell carcinoma lines was supported such that agents permissive for growth ranked as follows: Sea Plaque agarose approximately Sea Prep agarose greater than methyl cellulose greater than Bacto-agar. These observations have important implications for the in vitro study of transitional cell carcinoma cell lines and are relevant to the development of improved chemosensitivity determinations for human transitional cell carcinoma.

Topics: Agar; Carcinoma, Transitional Cell; Cell Division; Cell Line; Culture Media; Humans; In Vitro Techniques; Methylcellulose; Sepharose; Urinary Bladder; Urinary Bladder Neoplasms

To evaluate the persistence of cells with clonogenic properties in patients being treated for transitional cell carcinoma, 272 urine samples were collected from 75 patients and cultured in a double layer soft agar "cloning" system. The development of colonies was evaluated with growth curves based on repeated colony counting with an Omnicon automated colony counter at regular time intervals. Forty-eight patients had at least one evaluable culture. Comparing the results of colony development in culture with the clinical evaluation of the patients, 9 patients had a histologically proven recurrence preceded or accompanied by tumor colony growth in urine culture. One patient had tumor recurrence with growth negative urine culture (false negative). Fifteen patients have had growth negative urine culture with a negative follow-up (mean 19.5 months). Twenty-one patients have had growth positive urine cell cultures with no recurrence in their follow-up (mean 18.7 months). Although the follow-up times are at present relatively short, the present study suggests that repeated soft agar urine culture of patients with low-grade, low-stage bladder carcinoma may provide a means for identifying those patients at a higher risk for recurrence/progression of their disease.

Topics: Agar; Carcinoma, Transitional Cell; Cells, Cultured; Colony-Forming Units Assay; Follow-Up Studies; Humans; Neoplasm Recurrence, Local; Time Factors; Tumor Stem Cell Assay; Urinary Bladder Neoplasms; Urine

In this study we examined the ability of tumour specimens derived from bladder barbotage to produce cluster/colony formation in a tumour colony assay. In 114 bladder washings from 65 patients and 15 control subjects, we found that cluster and colony formation was highest from bladder washings obtained from patients with biopsy proven bladder cancer who were not on intravesical chemotherapy. Growth rates were extremely low, restricting the usefulness of the in vitro assay in its present form. This study suggests that improvements in the growth rates in the tumour colony assay will be necessary before this system can have real value in monitoring transitional cell carcinoma of the bladder.

Topics: Agar; Carcinoma, Transitional Cell; Clone Cells; Cytological Techniques; Humans; Urinary Bladder Neoplasms

Soft-agar cultures of transitional cell carcinoma from urine, irrigation fluid and transurethral resection solid tumor specimens show good colony growth. Growth of tumor colonies produced from urine was adequate to evaluate the presence of a viable tumor, since 12 of 18 noninfected cultures showed growth. The number of colonies produced was adequate to evaluate the sensitivity or resistance to chemotherapy agents in only 3 of 18 cultures. For irrigation fluid, similar results were obtained: 9 of 17 noninfected cultures showed growth, while only 3 of 17 were adequate for drug-sensitivity evaluation. For the evaluation of sensitivity or resistance, tumor cell suspensions were most appropriate, 7 of 25 noninfected cultures showed 30 or more colonies/dish, whereas 17 of 25 showed growth. With the possibility of obtaining growth of tumor cells derived from urine, a prospective study is proposed to define the value of repeated urine cultures in monitoring the status of the urothelium of patients treated for transitional cell carcinoma.

Topics: Agar; Biopsy; Carcinoma, Transitional Cell; Cells, Cultured; Cytological Techniques; Humans; Neoplasm Staging; Therapeutic Irrigation; Urinary Bladder; Urinary Bladder Neoplasms; Urine

Cellular growth interactions were studied between neonatal human lung fibroblasts (NLF-13) and human tumor lines derived from carcinomas of the prostate (PC-3, DU145), bladder (J82), and endometrium (HEC-1A) and from a glioma (Hs 683t). NLF-13 were interacted with tumor cells in soft agar or agarose media using two experimental protocols. In one system, NLF-13 cells were grown as anchored monolayers proliferating under the tumor cell layer. In the second, NLF-13 were embedded directly (nonanchored) into the agar or agarose layer with the tumor cells. The results from both interaction systems were similar for all five tumor lines. Anchored NLF-13 caused a dose-dependent inhibition of tumor growth, whereas nonanchored cells produced a dose-dependent growth stimulation. A time exposure experiment indicated that tumor stimulation and inhibition were biphasic responses to NLF-13. It was concluded that low concentrations of a diffusible NLF-13 product(s) accelerated tumor growth, whereas high concentrations were inhibitory. Further, the production of the active NLF-13 substance(s) was positively correlated with NLF-13 growth rate. Tumor cell inhibition was irreversible after a 5-day exposure to proliferating NLF-13 cells. Another line of normal neonatal human lung fibroblasts (NLF-147) showed inhibitory properties similar to those described for NLF-13. However, preliminary studies with fibroblasts from the skin of a Down's syndrome neonate (DS-172) and from a human kidney tumor (KTF-130) have shown both these fibroblast types to have a reduced ability to inhibit tumor cell cultures (J82) compared to the neonatal lung fibroblasts (NLF-13 and NLF-147).

Topics: Agar; Cell Communication; Cell Division; Cell Line; Female; Fibroblasts; Glioma; Humans; Infant, Newborn; Kinetics; Lung; Male; Prostatic Neoplasms; Urinary Bladder Neoplasms; Uterine Neoplasms

The effects of different agars (Bacto-agar and deoxycholate lactose agar), agaroses (LE, ME, Sea Plaque and Sea Prep 15/45) and methyl cellulose on the growth of a human tumor cell line, derived from a transitional cell carcinoma of the urinary bladder, were examined. The overall growth in the presence of agars and agarose was generally less than in liquid medium alone. In contrast, growth in the presence of methyl cellulose was significantly enhanced. Thus, methyl cellulose may be a useful agent for optimizing the proliferation of primary tissue cultures prepared from human transitional cell carcinomas.

Topics: Agar; Carcinoma, Transitional Cell; Cell Division; Cell Line; Culture Media; Humans; Methylcellulose; Sepharose; Urinary Bladder Neoplasms

Four human bladder carcinoma cell lines have been characterized by G-banding, and the chromosomal patterns correlated to growth in agar and tumorigenicity in nude mice. Each cell line was shown to be chromosomally unique and although numerical and structural anomalies were present, none were common to all four cell lines. However, one or more copies of a structurally altered chromosome 8 were present in all four cell lines and may be associated with tumorigenicity in nude mice. A combination of three marker chromosomes was found in the more anaplastic cell lines, but not in the two well-differentiated tumour cell lines. Growth in agar may be associated with the presence of the three marker chromosomes but was not correlated with tumorigenicity in nude mice.

Topics: Agar; Animals; Cell Line; Chromosome Banding; Chromosomes; Humans; Karyotyping; Mice; Mice, Nude; Neoplasm Transplantation; Transplantation, Heterologous; Urinary Bladder Neoplasms

Cytogenetic analysis of human tumors is rapidly coming of age as a useful tool in the diagnosis and prognosis of cancer. With the use of chromosome-banding analysis, an increasing display of tumors containing cytologically recognizable nonrandom chromosome change is accumulating. Additionally, with avenues of assessing drug resistance and sensitivity, current cytogenetic analyses are yielding clinically relevant information. Finally, the blending of cytogenetics with biochemistry and cell biology is yielding an impressive array of research tools for the subcellular study of cancer. DNA hybridization, gene cloning, autoradiography of various drug and hormonal chromosomal binding sites, gene mapping, and numerous other related techniques are providing fresh insights into the genetic basis of cancer. It seems clear that during the next decade substantial progress in understanding chromosome structure and function in normal cells as well as tumor cells will occur. Technical advances in the methodology for growing and harvesting tumor cells for detailed cytogenetic analysis will undoubtedly be important in this effort. Application of the described soft agar colony technique may provide a useful tool for study of the genetics of human solid tumors.

Topics: Agar; Chromosome Aberrations; Chromosome Disorders; Chromosomes, Human, 1-3; Clone Cells; Culture Media; Female; Humans; Karyotyping; Mitosis; Neoplasms; Ovarian Neoplasms; Urinary Bladder Neoplasms

Topics: Agar; Carcinoma; Cell Division; Cell Survival; Cells, Cultured; Clone Cells; Drug Evaluation, Preclinical; Humans; Karyotyping; Methylcellulose; Mitomycins; Urinary Bladder Neoplasms

We report the development of a clonogenic assay for progenitor cells in transitional cell carcinoma of the bladder. Colony growth has been demonstrated from cells obtained both from surgical biopsies and from bladder barbotages. Electron microscopic and karyotypic evidence supports the contention that these progenitors represent a part of the population maintaining the tumor in vivo. Colony growth occurred in 9 of 11 surgical biopsy samples and in 6 of 6 bladder barbotage samples. Plating efficiency ranged up to 0.7%, and colony size was in some instances greater than 1000 cells. The assay appears potentially useful for analysis of the biology of human transitional cell carcinoma.

Topics: Agar; Aged; Carcinoma, Transitional Cell; Cell Division; Chromosome Aberrations; Clone Cells; Female; Humans; Male; Methods; Methylcellulose; Microscopy, Electron; Middle Aged; Urinary Bladder Neoplasms

The frequency of M-components was studied by agar gel electrophoresis in sera from 807 patients, 467 (57%) females and 340 (43%) males with histologically proven solid malignant neoplasms.M-components were found in the sera of 40 male and 20 female patients. Apart from two known cases of multiple myeloma and one case of Waldenström's macroglobulinaemia, none of the patients were found to be suffering from these diseases. The frequency of M-components increased with age, and this was more evident in males. Twenty-two of 60 patients with M-components did not exhibit abnormalities on immunoelectrophoresis. Of the 35 remaining patients, 27 had an abnormal component of the IgG class, 6 of the IgA and 2 of the IgM class. M-components were found in the sera of patients with a wide variety of neoplasms. There appeared to be no evidence of an increased frequency of M-components in the sera of patients with solid malignant neoplasms compared with normal adult population.

Topics: Adolescent; Adult; Agar; Aged; Child; Electrophoresis; Female; Gastrointestinal Neoplasms; Genital Neoplasms, Female; Humans; Immunoelectrophoresis; Immunoglobulin A; Immunoglobulin G; Immunoglobulin M; Immunoglobulins; Lung Neoplasms; Male; Middle Aged; Multiple Myeloma; Neoplasms; Urinary Bladder Neoplasms; Waldenstrom Macroglobulinemia

Topics: Agar; Aniline Compounds; Animals; Carcinogens; Carcinosarcoma; Escherichia coli; Genetics, Microbial; Injections, Intraperitoneal; Leukemia, Experimental; Male; Mutagens; Mutation; Naphthalenes; Naphthoquinones; Neoplasms, Experimental; Peritoneal Neoplasms; Peritoneum; Rats; Sarcoma, Experimental; Urinary Bladder Neoplasms