agar and Carcinoma

Topics: Agar; Amino Acid Sequence; Animals; Buffers; Carcinoma; Cathepsins; Chemical Phenomena; Chemistry; Chromium Isotopes; Dietary Proteins; Duodenum; Electrophoresis; Enzyme Precursors; Gastric Juice; Gels; Humans; Hydrochloric Acid; Hydrogen-Ion Concentration; Molecular Weight; Pepsin A; Stomach; Stomach Neoplasms; Stomach Ulcer; Swine; Terminology as Topic

Oral squamous cell carcinoma (OSCC) is the most common type of oral cancer. Despite progress in the treatment of OSCC, overall survival has not improved substantially in the last three decades. Therefore, identification of reliable biomarkers becomes essential to develop effective anti-cancer therapy. In this study, we focused on the enzyme Nicotinamide N-methyltransferase (NNMT), which plays a fundamental role in the biotransformation of many xenobiotics. Although several tumors have been associated with abnormal NNMT expression, its role in cancer cell metabolism remains largely unknown. In this report, 7 human oral cancer cell lines were examined for NNMT expression by Real-Time PCR, Western blot and HPLC-based catalytic assay. Subsequently, we evaluated the in vitro effect of shRNA-mediated silencing of NNMT on cell proliferation. In vivo tumorigenicity of oral cancer cells with stable knockdown of NNMT was assayed by using xenograft models. High expression levels of NNMT were found in PE/CA PJ-15 cells, in keeping with the results of Western blot and catalytic activity assay. PE/CA PJ-15 cell line was stably transfected with shRNA plasmids against NNMT and analyzed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) and soft agar Assays. Transfected and control cells were injected into athymic mice in order to evaluate the effect of NNMT silencing on tumor growth. NNMT downregulation resulted in decreased cell proliferation and colony formation ability on soft agar. In athymic mice, NNMT silencing induced a marked reduction in tumour volume. Our results show that the downregulation of NNMT expression in human oral carcinoma cells significantly inhibits cell growth in vitro and tumorigenicity in vivo. All these experimental data seem to suggest that NNMT plays a critical role in the proliferation and tumorigenic capacity of oral cancer cells, and its inhibition could represent a potential molecular approach to the treatment of oral carcinoma.

Topics: Agar; Animals; Biomarkers, Tumor; Blotting, Western; Carcinoma; Cell Line, Tumor; Cell Proliferation; Chromatography, High Pressure Liquid; Female; Gene Silencing; Humans; Male; Mice; Mice, Inbred BALB C; Mice, Nude; Mouth Neoplasms; Neoplasm Transplantation; Nicotinamide N-Methyltransferase; Real-Time Polymerase Chain Reaction; RNA Interference; RNA, Small Interfering; Tetrazolium Salts; Thiazoles

The cyclin-dependent kinase inhibitor p21cip1/waf1 negatively regulates the progression of cell cycle and the potential usefulness of p21cip1/waf1 gene is proposed in gene therapy. However, studies have demonstrated a protective role of p21cip1/waf1 against apoptosis and little is known about effects of ectopic expression of p21cip1/waf1 on differentiation of colon cancer cells. In the present study, we found diffuse p21cip1/waf1 expression in only a few clinical samples of colorectal cancer with wild-type p53 gene. To explore the role of p21cip1/waf1 in cell growth, apoptosis and differentiation, we constitutively overexpressed p21cip1/waf1 in HT29 colon carcinoma cells. Ectopic overexpression of p21cip1/waf1 was associated with inhibition of CDK2-associated kinase activity, indicating the functionality of the introduced p21cip1/waf1 gene. Overexpression of p21cip1/waf1 caused an appreciable growth inhibition in monolayer and soft agar cultures and it significantly reduced sodium butyrate- but not 5-fluorouracil-induced apoptosis. p21cip1/waf1 overexpressing cells exhibited marked decrease of intestinal differentiation when assayed with intestinal alkaline phosphatase. Our findings suggest that introduction of p21cip1/waf1 gene into colon cancer cells may be useful for inhibiting cell growth but caution should be taken regarding the increased resistance to certain apoptosis-inducing agents and dysregulation of endogenous p21cip1/waf1-mediated differentiation process.

Topics: Agar; Alkaline Phosphatase; Apoptosis; Blotting, Western; Butyrates; Carcinoma; Cell Cycle Proteins; Cell Differentiation; Cell Line, Tumor; Cell Proliferation; Colonic Neoplasms; Cyclin-Dependent Kinase Inhibitor p21; DNA Mutational Analysis; Flow Cytometry; Fluorouracil; Gene Expression Regulation, Neoplastic; Humans; Inhibitory Concentration 50; Isobutyrates; Mutation; Polymerase Chain Reaction; Polymorphism, Single-Stranded Conformational; Time Factors; Tumor Suppressor Protein p53

Undifferentiated (anaplastic) thyroid carcinoma is a highly aggressive human cancer with very poor prognosis. Although there have been a few studies of candidate treatments, the fact that it is an infrequent tumor makes it very difficult to design clinical trials. A strong association has been observed between undifferentiated thyroid carcinoma and TP53 mutations in numerous molecular genetic and expression studies. Plitidepsin (Aplidin, PharmaMar, Madrid, Spain) is a novel anticancer compound obtained from a sea tunicate. This compound has been reported to induce apoptosis independently of TP53 status. We investigated the actions of plitidepsin in human thyroid cancer cells. In initial experiments using primary cultured cells from a differentiated (papillary) carcinoma, we found that 100 nmol/L plitidepsin induced apoptosis, whereas lower doses were cytostatic. Because our aim was to study the effects of plitidepsin at clinically relevant concentrations, subsequent experiments were done with a dosage regimen reflecting plasma concentrations observed in previously reported clinical trials: 100 nmol/L for 4 hours, followed by 10 nmol/L for 20 hours (4(100)/20(10) plitidepsin). This plitidepsin dosage regimen blocked the proliferation of a primary undifferentiated/anaplastic thyroid carcinoma culture obtained in our laboratory and of a commercial cell line (8305C) obtained from an undifferentiated thyroid carcinoma; however, it did not induce apoptosis. The proportion of cells in the G(1) phase of the cell cycle was greatly increased and the proportion in the S/G(2)-M phases greatly reduced, suggesting that plitidepsin blocks G(1)-to-S transition. Levels of the cyclin D1/cyclin-dependent kinase 4/p21 complex proteins were decreased and, in line with this, the levels of unphosphorylated Rb1 increased. The decrease in cell cycle proteins correlated with hypoacetylation of histone H3. Finally, we did experiments to assess how rapidly tumor cells return to their initial pretreatment proliferative behavior after 4(100)/20(10) plitidepsin treatment. Cells from undifferentiated tumors needed more than 3 days to recover logarithmic growth, and after 7 days, cell number was still significantly lower than in control cultures. 4(100)/20(10) plitidepsin inhibited the growth in soft agar. Together, our data show that plitidepsin is able to block in vitro cell cycle progression at concentrations similar to serum concentrations observed in vivo, and that this effect is pe

Topics: Adult; Agar; Aged; Antineoplastic Agents; Apoptosis; Carcinoma; Cell Cycle; Cell Differentiation; Cell Line, Tumor; Cell Proliferation; Clinical Trials as Topic; Depsipeptides; Dose-Response Relationship, Drug; Female; Flow Cytometry; Gene Expression Regulation, Neoplastic; Genes, p53; HeLa Cells; Histones; Humans; Immunoblotting; Male; Middle Aged; Models, Statistical; Peptides, Cyclic; Thyroid Gland; Thyroid Neoplasms; Time Factors; Tumor Cells, Cultured; Tumor Suppressor Protein p53

Carcinoma-associated fibroblasts (CAF) promote tumor progression of pre-neoplastic epithelial cells. To investigate the basis of this phenomenon, we compared the properties of fibroblasts cultured from normal human prostate (NHPF) to prostate CAF. NHPF and CAF were assayed for growth potential, cell death and proliferative capacity by measuring population doubling time, cell cycle distribution and capability to form colonies in soft agar. Resistance to genotoxic (UV radiation: 0-50 J/cm2) and chemotoxic (0-200 nM Taxol) agents were compared between CAF and NHPF by measuring cell viability and cell cycle analysis. Transforming growth factor beta1 (TGF-beta1) immunoreactivity was assessed in non-malignant and malignant prostatic tissue. No detectable differences were found when comparing CAF and NHPF with respect to population doubling time, cell cycle distribution and response to genotoxic and chemotoxic agents. The mean number of colonies in soft agar was 120.5 for CAF vs. 18.2 for NHPF (p < 0.05). Because TGF-beta1 and matrix metalloproteinase (MMP)-9 have been associated with growth of fibroblasts in soft agar and tumor promotion, we measured the expression of these factors in NHPF and CAF by ELISA. There was no difference in expression of MMP-9; however, TGF-beta1 was expressed in higher concentrations in CAF than in NHPF (p < 0.0014). Furthermore, TGF-beta1 expression was higher in the carcinoma-associated stroma of prostate cancer tissue than stroma of non-malignant prostatic tissue. Increased capability of CAF as compared to NHPF to form colonies in soft agar may be due to a higher expression of TGF-beta1 and correlates with the ability of CAF to promote malignant progression of prostate epithelial cells.

Topics: Agar; Antineoplastic Agents; Carcinoma; Cell Cycle; Cell Death; Cell Division; Culture Media; Drug Resistance, Neoplasm; Enzyme-Linked Immunosorbent Assay; Fibroblasts; Gene Expression Profiling; Humans; Male; Prostate; Prostatic Neoplasms; Radiation Tolerance; Transforming Growth Factor beta; Transforming Growth Factor beta1; Tumor Cells, Cultured

To determine the thermal dose of a hyperthermia treatment, knowledge of the three-dimensional (3D) temperature distribution is mandatory. The aim of this paper is to validate an interstitial hyperthermia treatment planning system with which the full 3D temperature distribution can be obtained in individual patients. Within a phase I study, 12 patients with prostate cancer were treated with interstitial hyperthermia using our multi electrode current source interstitial hyperthermia treatment (MECS IHT) system. The temperature distribution was measured from within the heating devices and by additional thermometry. The perfusion level was estimated and the heating implant reconstructed. The steady-state temperature distribution was calculated using our interstitial hyperthermia treatment planning system. The simulated temperature distribution was validated by individually comparing the measured and simulated thermo-sensors, both for the thermometry integrated with the heating applicators and the additional thermometry. The entire procedure was also performed on a no-flow agar-agar phantom. It was shown that the calculated temperature distribution of an individual patient during MECS interstitial hyperthermia is very heterogeneous. The validation indicates that the calculated temperature elevations match the measurements within approximately 1 degrees C. Possible improvements are more precise reconstruction, incorporation of discrete vasculature and using a temperature-dependent, heterogeneous perfusion distribution. Further technical improvements of the MECS-IHT system may also result in better temperature calculations.

Topics: Agar; Calibration; Carcinoma; Electrodes; Humans; Hyperthermia, Induced; Male; Phantoms, Imaging; Prostatic Neoplasms; Reproducibility of Results; Temperature; Time Factors

A technique for agar embedding of bone marrow aspirate particles is compared with the conventional aspirate smear and bone marrow biopsy by reviewing 503 consecutive bone marrow specimens. Immunohistochemical studies were performed on both agar sections and bone marrow biopsies on 43 paired specimens to compare the results between the two preparations. The results were also compared with traditional clot sections from ten control cases. Of the 382 cases with agar sections, 97.7% contained material in the agar that was diagnostic alone or supportive of the diagnosis made with the biopsy and aspirate smear. In two cases (0.4%), focal involvement by lymphoma was identified on the agar section but not in the biopsy sections or aspirate smears. The immunohistochemical studies showed superior immunoreactivity in agar sections by lymphoproliferative disorders when compared with bone marrow biopsy sections. Similar results between agar and conventional biopsy sections were found in cases of metastatic carcinoma and plasma cell dyscrasias.

Topics: Adolescent; Adult; Agar; Aged; Aged, 80 and over; Biopsy; Bone Marrow; Carcinoma; Cell Aggregation; Child; Child, Preschool; Female; Histological Techniques; Humans; Immunohistochemistry; Infant; Leukemia, Prolymphocytic, T-Cell; Lymphocytes; Lymphoproliferative Disorders; Male; Middle Aged; Paraproteinemias

Concurrent Cloning Efficiencies (CE) of both ascites and solid tumor samples from 36 patients with ovarian carcinoma were studied using the soft agar assay. The CE of both were highly variable (range, 0-1.234% and 0-0.802%, respectively). There was marked intrapatient and interpatient heterogeneity in the CE. Of the 36 tested, comparative CE were evaluable in 29. CE was 0 in both solid tumor and ascites in one patient. CE was 0 in four other ascites samples from four patients. In other 24, the relative CE of solid tumor/ascites from each patient ranged from 0.066 to 435. In the 29 patients with samples of ascites and a solid tumor evaluable for concurrent CE, the median colony counts of solid tumors was more than tenfold higher than ascites. The solid tumors obtained from 31 patients had a significantly higher CE than tumor cells obtained from ascites samples from 32 patients. Solid tumors were significantly better than ascites for in vitro testing based on the data that 75% (27/36) of solid tumors and only 31% (11/36) of ascites formed greater than or equal to 30 colonies. The drug sensitivity profiles of tumor cells from a solid tumor and ascites of the same patient appear similar. Based on these observations, it may be more cost and labor effective to do soft agar in vitro chemotherapy assays using a solid tumor than ascites in ovarian carcinoma.

Topics: Agar; Ascites; Carcinoma; Colony-Forming Units Assay; Drug Screening Assays, Antitumor; Female; Humans; Ovarian Neoplasms; Tumor Stem Cell Assay

We studied the factors that control the capacity of tumor cells to grow in soft agar. And, we analyzed the effect of cell-free ascites (CFA) obtained from ovarian cancer patients in combination with various media on the cloning efficiency of H-134 and OVCAR-3nu ovarian carcinoma cell lines and the WiDr colon carcinoma cell line. Seven batches of CFA consistently enhanced the soft agar growth of tumor cells more efficiently than tested sera. The addition of charcoal-treated bovine serum albumin (BSA) lowered the amount of CFA required for optimal tumor cell growth. As little as 1.25 ng of epidermal growth factor (EGF)/ml further improved OVCAR-3nu soft agar growth in combination with all of the amounts of CFA tested. Thus, neither BSA nor EGF seems to account for the colony-stimulating effect of ascites on tumor cells. Four batches of CFA were tested for stimulating soft agar growth of normal rat kidney (NRK-49F) fibroblasts; all induced colonies of different morphologies. This effect was potentiated by EGF, which suggests the presence of several transforming growth factor-like activities in CFA. The results show that differences in cloning efficiency of tumor cells of one or two orders of magnitude can be found between standard (anchorage-dependent growth-supporting) media and media optimalized for soft agar growth, such as CFA in the presence of EGF. This paper will discuss the similarity in effects of CFA on various tumor cells and NRK cells, and possible implications of the stimulatory effects of CFA.

Topics: Agar; Animals; Ascites; Carcinoma; Cell Division; Cells, Cultured; Clone Cells; Colonic Neoplasms; Female; Humans; Neoplastic Stem Cells; Ovarian Neoplasms; Rats

The human tumor clonogenic assay (HTCA) has potential value for studies of both the chemosensitivity and biology of human tumors. However, many technical problems including low plating efficiencies and the preparation of sufficient numbers of viable cells remain. In this study, an improved method for disaggregation of solid tumors increased the yield of single cells. Consequently, more than 10 anticancer drugs could be tested in 94 of 168 specimens (56%). Removal of peripheral blood lymphocytes from cell suspensions derived from effusions also improved colony formation. Adequate growth for sensitivity testing (greater than 30 colonies/plate) was obtained in 122 cases (73%), inadequate growth for drug evaluation (5-29 colonies/plate) in 29 cases (17%), and no colony formation (less than 5 colonies/plate) in 17 cases (10%) of the 168 viable samples. The cloning efficiencies of cells derived from primary tumors (median 0.015%) were higher than those of cells derived from metastatic tumors (0.012%), and they varied with the location of the metastatic site. Cloning efficiencies varied markedly from specimen to specimen, and were unaffected by tumor histology, grade of differentiation, patient age, stage of disease, or prior chemotherapy. The HTCA is promising as a potential tool for studying the biology of tumors.

Topics: Agar; Carcinoma; Cell Aggregation; Colony-Forming Units Assay; Humans; Lung Neoplasms; Neoplasm Staging; Prognosis; Tumor Stem Cell Assay

Thirty-one bone marrow aspirations were performed on patients with prostatic carcinoma metastatic to bone. After separation over a Ficoll-Hypaque gradient viable nucleated cells were cultured in semisolid agar. Colony formation occurred in 14 of 27 (52%) nonbacterially contaminated cultures. Characterization of cells from the colonies showed them to be consistent with malignant prostate cells. After staining, these cells were periodic acid-Schiff positive, prostatic acid phosphatase positive, and prostatic specific antigen positive. Other studies demonstrated the cells to be karyotypically abnormal, ultrastructurally similar to epithelial cells, and capable of secondary colony formation. Three bone marrow aspirate specimens did not have metastatic prostatic carcinoma detected by standard methods but did demonstrate colony formation. However, colony formation was most frequently seen when a radionuclide scan was positive at the aspiration site and when tumor cells were microscopically detectable by Wright staining of a smeared aspirate. The potential utility of colony forming cultures in prostate cancer is discussed. In working with bone marrow aspirates, additional cell separation procedures may be required to calculate and maximize plating efficiencies.

Topics: Agar; Biopsy; Bone Marrow; Bone Neoplasms; Carcinoma; Cells, Cultured; Humans; Karyotyping; Male; Microscopy, Electron; Neoplasm Metastasis; Prostatic Neoplasms

A continuous human cell line was established from a patient with large cell anaplastic lung carcinoma. This cell line, designated QU-DB, has been in culture for over 36 months and grows as an adherent monolayer with a doubling time of 10-12 hours. Its morphology, ultrastructure, karyotype, ability to grow in soft agar and heterotransplantability, indicate it is a large-cell lung tumor cell line of human origin. Three cell lines were established from metastatic tumors in nude mice receiving subcutaneous injections of QU-DB cells. The morphology and growth characteristics exhibited by these cell lines were similar to the primary cell line. Karyotypic analysis of cell lines derived from the primary tumor and a metastasis to the diaphragm were similar, but cells from a liver metastasis culture showed additional karyotypic changes. This large cell lung tumor cell line may prove useful as a model system for studies of human tumor progression and metastasis.

Topics: Agar; Aged; Animals; Carcinoma; Cell Division; Cell Line; Clone Cells; Flow Cytometry; Humans; Karyotyping; Liver Neoplasms; Lung Neoplasms; Male; Mice; Mice, Nude; Microscopy, Electron; Neoplasm Metastasis; Smoking

In C57Bl/6 mice with different stages of Lewis lung carcinoma growth used as recipients of diffusion chambers, after subcutaneous transplantation of tumour cells there was an increased output of colonies in the bone marrow and spleen cell cultures (CFU-DC) as compared to the control, The cocultivation of normal (bone marrow and spleen) and tumour cells has no influence on the effectiveness of their cloning.

Topics: Abdomen; Agar; Animals; Bone Marrow; Bone Marrow Cells; Carcinoma; Cell Count; Cells, Cultured; Diffusion; Hematopoietic Stem Cells; Lung Neoplasms; Mice; Mice, Inbred C57BL; Spleen; Time Factors

Four families of human tumour cell lines--one of uterine, and three of ovarian origin--were established at early passage level from primary biopsy specimens of terminal patients by the propagation of anchorage-independent agar clones in liquid culture. All cell lines exhibited unique and stable characteristics and retained their ability to clone in agar. However, considerable heterogeneity was evident in clonogenic capacity, karyotype, and responsiveness to growth-promoting substances even among progeny of single agar clones isolated from the one biopsy specimen. These cell lines will be made available for further study upon request.

Topics: Agar; Aged; Biopsy; Carcinoma; Cell Line; Culture Media; Cystadenocarcinoma; Cytological Techniques; Female; Humans; Middle Aged; Neoplasms; Neoplasms, Experimental; Ovarian Neoplasms; Uterine Neoplasms

Methods and evaluation of the human tumor stem cell assay (HTSCA) are described. Advantages and disadvantages of the test system are elaborated. The in vitro/in vivo correlation in the drug screening of human ovarian carcinomas shows that the prediction of sensitivity to a cytotoxic agent is only possible in 64%. Prediction of drug resistance, however, seems to be possible in 95%. The number of patients that profit from the HTSCA seems to be only less than 10%. Our investigations describe the influence of various hormones and antiestrogens on the colony formation of human ovarian carcinoma cells. Tamoxifen and his major metabolite 4-hydroxy-tamoxifen were the most active agents. Both compounds inhibit the colony survival (70% at pharmacological concentrations) of 60% of the screened ovarian carcinomas. A significant correlation to the quantitative level of estrogen or progesterone receptors could not be proved. Colony formation of ovarian carcinoma cells was compared in the HTSCA as described by Hamburger and Salmon and in a methylcellulose-monolayer system. Our results show that the colony formation corresponds to the results of the original HTSCA: Cloning ovarian carcinoma cells in the methylcellulose-monolayer, however, seems to be technically easier and faster.

Topics: Agar; Antineoplastic Agents; Carcinoma; Cells, Cultured; Colony-Forming Units Assay; Drug Resistance; Estradiol; Female; Hormones; Humans; Medroxyprogesterone; Medroxyprogesterone Acetate; Methylcellulose; Ovarian Neoplasms; Progesterone; Prognosis; Tamoxifen; Tumor Stem Cell Assay

An in vitro soft agar culture system was utilized to evaluate the colony growth of cells from primary breast carcinoma. A total of 53 specimens from fifty-three patients were placed in culture. Of these, 29 samples (55 per cent) formed at least 30 colonies per 500,000 cells plated. In relation to histologic type of tumor and clinical status of the disease, 4 of 4 samples from mucous carcinoma grew into colonies and, then, t-categories, i.e. histological extent of primary tumor, and colony growth showed an inverse correlation. Estrogen receptor status did not appear to influence growth of the colonies. The in vitro sensitivity studies to adriamycin showed a dose dependent increase in lethality. However, the in vitro response rate was relatively low. This assay system can be used to study the biology and clinical approaches to treatment of breast carcinoma.

Topics: Adult; Agar; Aged; Breast Neoplasms; Carcinoma; Cell Division; Cell Survival; Cells, Cultured; Colony-Forming Units Assay; Doxorubicin; Female; Humans; Middle Aged; Receptors, Estrogen; Tumor Stem Cell Assay

A previously unpublished reaction of precipitation in agarose between histone and non-histone proteins extracted in saline buffer from nuclei of human skin tumors, is reported. Two bands of precipitation similar to those in an immunodiffusion reaction between NHP and specific antisera, were observed. The reaction described is very similar to the affinodiffusion reaction of glycoproteins and lectins in agarose.

Topics: Agar; Animals; Carcinoma; Chromosomal Proteins, Non-Histone; Fibroma; Histones; Humans; Immunodiffusion; Melanoma; Rats; Rats, Inbred Strains; Skin Neoplasms

Human carcinoma tissues were grown in culture for two to four weeks using the two-layer soft agar technique. All cultures that showed growth of tumor cell colonies also showed well-preserved, apparently healthy tumor cells lying singly. These cells showed neither proliferative capacity nor necrosis or morphologic degeneration during the time in soft agar. Thus, morphologic criteria seem to be poor indicators of tumor cell proliferative potential, at least in the short term. However, the method of soft agar tumor clonogenic cell culture itself provided a direct measure of tumor cell proliferative capacity, ie, the formation of colonies from single tumor cells. This may be valuable in directly assessing the presence of "viable" tumor cells in biopsy specimens taken after therapy, and thus guide further patient therapy.

Topics: Agar; Aged; Carcinoma; Cell Transformation, Neoplastic; Cells, Cultured; Female; Humans; Male

In vitro growth of tumor cells may provide a way of testing the effectiveness of chemotherapeutic agents in the treatment of cancer. In 1980 and 1981, operations for gynecologic malignancy were performed on 610 Mayo Clinic patients, and malignant tissue and fluids were obtained from 204 cancers that involved the vulva, uterus, fallopian tubes, and ovaries. These yielded 76 clonogenic stem-cell preparations; and various chemotherapeutic agents were tested against these 76 tumors on soft agar. Considered in this study were the overall process of culturing the samples of tumors and, especially, the data from the preparations that showed sufficient growth of tumor cells for testing. Our guiding concerns were the usefulness of this method to gynecologists and the possible benefits to patients.

Topics: Agar; Antineoplastic Agents; Carcinoma; Clone Cells; Culture Media; Drug Resistance; Female; Genital Neoplasms, Female; Humans; In Vitro Techniques; Ovarian Neoplasms

The effects of feeder layers of C3H/10T 1/2 cells on the growth of human and mouse cell lines were tested. Feeder layers increased colony formation by cultured cancerous cells in semisolid medium over controls grown in semisolid medium without feeder layers. Maximal colony formation was also attained at a faster rate when feeder layers were used. Plating efficiency by cancerous cells obtained directly from xenografts was increased threefold to fivefold in tissue culture when feeder cells were present as confluent monolayers.

Topics: Agar; Animals; Carcinoma; Cell Count; Cell Line; Colonic Neoplasms; Embryo, Mammalian; Humans; Mice; Mice, Inbred C3H; Neoplasm Transplantation; Time Factors

We studied the in vitro growth characteristics of 10 solid-tumor samples of patients with ovarian carcinoma using a semisolid agar culture technique. Tumor cell colonies were observed in 8 of 10 samples, but sufficient number of tumor colonies to evaluate the effects of interferon and other antitumor agents were obtained in only four samples. As compared with cell suspensions prepared from ascitic fluid samples, solid-tumor samples had markedly lower viability, 39% vs 89%, and had more tumor cells, 81% vs 28%. Also, whereas the maximum increase in tumor-colony number occurred during the first week of growth in both solid- and ascitic-fluid-derived samples grown concurrently from the same donors, increase in tumor colony number was sustained for longer periods in ascitic-fluid-derived cultures. The ascitic-fluid-derived tumor colonies were more sensitive to the antiproliferative effects of interferon than colonies derived from solid-tumors. At a concentration of 300 units/ml incorporated into the agar for the duration of the culture, three of four ascitic fluid samples showed a reduction in tumor colony number by greater than or equal to 25%, whereas none of the solid-tumor samples were affected by the interferon to that degree. In contrast, solid-tumor samples showed greater response to cis platinum and Adriamycin than did ascitic-fluid-derived cultures. Such studies and observations are critical in designing clinical trials for the use of interferon in the treatment of malignancy and the judicious selection of patients and route of administration most likely to provide optimal results, especially in view of present critical shortages in availability of interferon.

Topics: Adult; Agar; Aged; Ascites; Carcinoma; Cisplatin; Clone Cells; Culture Media; Doxorubicin; Female; Humans; Interferons; Middle Aged; Ovarian Neoplasms

The effect of mechanical and enzymatic disaggregation on human malignant melanoma, soft-tissue sarcoma and lung carcinoma colony growth in soft agar was studied. The enzymatic disaggregation was advantageous in most cases of melanoma and sarcoma, giving a larger number of colonies and increasing the probability of achieving growth in soft agar. Enzymatically treated pulmonary carcinoma cell populations had lower clonogeneic potential, especially in the case of anaplastic carcinomas. Morphological studies showed that the cells growing in soft-agar colonies had the same characteristics as those of the original tumor. A linear relationship was obtained between the number of enzymatically and mechanically treated tumor cells plated and the number of colonies. Delayed plating decreased the number of colonies.

Topics: Agar; Carcinoma; Cell Aggregation; Clone Cells; Cytological Techniques; Humans; Lung Neoplasms; Melanoma; Microbial Collagenase; Sarcoma; Soft Tissue Neoplasms; Time Factors

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

Various tissues and cells in culture contain a specific inhibitor of DNase I (EC 3.1.4.5). In this paper evidence is presented that this inhibitor is actin, one of the major structural proteins of muscle and nonmuscle cells. (a) The inhibitor is a major cellular component constituting 5-10% of the soluble protein. (b) It migrates with actin on sodium dodecyl sulfate-polyacrylamide gel electrophoresis, having a characteristic molecular weight of 42,000. (c) It has an amino-acid composition closely similar to that of actin. (d) The peptide maps of the two proteins are nearly identical. (e) Skeletal muscle actin inhibits the enzymatic activity of DNase I. (f) DNase I-agarose affinity chromatography quantitatively retains purified skeletal muscle actin, and actin, specifically, from high-speed supernatants of whole cell extracts. (g) An antibody to purified inhibitor protein from calf thymus, used in indirect immunofluorescence on cells grown in culture, stains a two-dimensional network of fibers similar to that seen with an actin-specific antibody.The observation that actin can be isolated by DNase-agarose affinity chromatography provides a useful tool for the biochemical study of actin under different physiological conditions.

Topics: Actins; Agar; Amino Acid Sequence; Animals; Carcinoma; Cattle; Cell Line; Chickens; Chromatography, Affinity; Deoxyribonucleases; Electrophoresis, Polyacrylamide Gel; Epitopes; Fibroblasts; Fluorescent Antibody Technique; Humans; Hydrolysis; Immunoassay; Molecular Weight; Mouth Neoplasms; Muscles; Peptide Fragments; Polysaccharides; Spleen; Thymus Gland; Trypsin

Areas of cytopathic effect can be circumscribed in cell monolayers by adding antiserum to the liquid nutrient medium after adsorption of virus. This procedure represents a simple and reliable tool for the titration of virus infectivity and provides an experimental model for studying some aspects of virus infection.

Topics: Adsorption; Agar; Animals; Carcinoma; Cell Line; Chick Embryo; Culture Techniques; Cytopathogenic Effect, Viral; Evaluation Studies as Topic; Haplorhini; Humans; Immune Sera; Kidney; Methods; Mouth Neoplasms; Orthomyxoviridae; Poliovirus; Rabbits; Viral Plaque Assay

Topics: Adenocarcinoma; Adenocarcinoma, Scirrhous; Adolescent; Adult; Agar; Aged; Carcinoma; Chromatography; Colonic Neoplasms; Gallbladder Neoplasms; Hemangiosarcoma; Humans; Liver Neoplasms; Lymphoma, Non-Hodgkin; Middle Aged; Neoplasm Proteins; Neoplasm Recurrence, Local; Pancreatic Neoplasms; Protein Denaturation; Retroperitoneal Neoplasms; Stomach Neoplasms

Epithelial-like cells originating from 10-day and 8-week old BD rats were treated in vitro with dimethylnitrosamine (DMN) and N-methyl-N'-nitro-nitrosoguanidine (MNNG). Morphologically, no differences were seen between treated and untreated cells up to the time when the cells were transplanted into syngeneic hosts. However, the treated cells grew in soft agar and once injected subcutaneously or intraperitoneally into newborn rats, developed tumours after a latent period of 9-12 weeks. The tumours obtained with DMN-treated cells were well differentiated adenocarcinomata, whereas carcinosarcomata were observed with the MNNG-treated cells.

Topics: Adenocarcinoma; Agar; Animals; Carcinoma; Cell Line; Epithelial Cells; Injections, Intraperitoneal; Injections, Subcutaneous; Liver Neoplasms; Neoplasm Transplantation; Nitrosamines; Nitrosoguanidines; Rats

Topics: Agar; Animals; Antineoplastic Agents; Carcinoma; Cell Line; Cells, Cultured; Disease Models, Animal; Humans; Leukemia L1210; Melanoma; Mice; Mice, Inbred Strains; Mouth Neoplasms; Neoplasm Transplantation; Neoplasms, Experimental; Oncogenic Viruses; Plant Extracts

Topics: Agar; Carcinoma; Electrophoresis; Gels; Half-Life; Hodgkin Disease; Humans; Isoenzymes; L-Lactate Dehydrogenase; Leukemia, Myeloid; Leukemia, Myeloid, Acute; Leukocytes; Lymphatic Diseases; Lymphoma; Neoplasms; Osteosclerosis; Plasmacytoma; Sarcoma; Tissue Extracts

Topics: Agar; Breast Diseases; Breast Neoplasms; Carcinoma; Computers; Electrophoresis; Isoenzymes; L-Lactate Dehydrogenase

Topics: Agar; Animals; Carcinoma; Carcinoma, Hepatocellular; Chemical Precipitation; Cricetinae; Electrophoresis; gamma-Globulins; In Vitro Techniques; Kidney; Liver Neoplasms; Neoplasm Proteins; Neoplasms, Experimental; Proteins; Quaternary Ammonium Compounds; Rats; Tissue Extracts

Topics: Agar; Animals; Antineoplastic Agents; Carcinoma; Carcinoma, Ehrlich Tumor; Chemistry, Pharmaceutical; Enzyme Inhibitors; Mice; Oxidoreductases; Pharmacology; Research; Sarcoma 180; Sarcoma, Yoshida

Topics: Agar; Biophysical Phenomena; Carcinoma; Humans; Neoplasms

Topics: Agar; Carcinoma; Early Diagnosis; Female; Humans; Uterine Cervical Neoplasms