Clonogenic cell assay for anchorage‐dependent squamous carcinoma cell lines using limiting dilution

Grénman, Reidar; Burk, David W.; Virolainen, Erkki; Buick, Roger; Church, Jon G.; Schwartz, Donald R.; Carey, Thomas E.

doi:10.1002/ijc.2910440123

Cited by 77 publications

(47 citation statements)

References 26 publications

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“…F4 and F5 are the cell clones isolated from the SOSP-9607 cell line using the limited dilution method (Grenman et al 1989). All the cell lines were maintained in complete RPMI 1640 medium (HyClone) supplemented with 10% fetal calf serum (FCS) (Sijiqing Co.) at 37°C under 5% CO 2 atmosphere.…”

Section: Cell Linesmentioning

confidence: 99%

Establishment and characterization of human osteosarcoma cell lines with different pulmonary metastatic potentials

et al. 2009

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To develop an investigative tool for the study of human osteosarcoma (OSA), we established a human OSA cell line, namely, SOSP-9607, which exhibits a potential for spontaneous pulmonary metastasis. Subsequently, we screened two related sublines (F5M2 and F4) that have different pulmonary metastatic potentials. An in vivo orthotopic transplantation assay confirmed spontaneous pulmonary metastasis in all mice (100%) transplanted with the more aggressive OSA cells (F5M2) and a lesser degree of metastases with smaller nodules in 33.3% mice transplanted with the less aggressive OSA cell subline (F4). In mice transplanted with F5M2 cells, death from metastasis occurred at a median of 71 days; however, in mice transplanted with F4, no death occurred even after 120 days. Therefore, the F5M2 and F4 sublines, which originated from the same parent cell line, differed with respect to metastasis-related properties such as proliferating ability and invasiveness. Hence, these well-characterized human OSA sublines can be used as valuable models for comparative studies of genetic determinants of OSA in the future.

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Section: Cell Linesmentioning

confidence: 99%

Establishment and characterization of human osteosarcoma cell lines with different pulmonary metastatic potentials

et al. 2009

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“…The clonogenic assay was performed as described previously. 27,28 Briefly, the cells were harvested with trypsin and EDTA, counted and diluted to a stock solution of 4,167 cells/ml. The number of cells plated per well was adjusted according to the plating efficiency (PE) of each cell line.…”

Section: Clonogenic Assay and Irradiationmentioning

confidence: 99%

Paclitaxel combined with fractionated radiation in vitro: A study with vulvar squamous cell carcinoma cell lines

Raitanen

Rantanen

Kulmala

et al. 2001

Intl Journal of Cancer

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Concurrent paclitaxel and radiation has given promising results in the treatment of a variety of solid tumors. We wanted to test the efficacy of this combination for vulvar carcinoma, which currently has a poor outcome in advanced stages. The radiation sensitivity, sublethal damage repair (SLDR) capacity and effect of paclitaxel during fractionated radiation were assessed in our study on 7 vulvar inherently radioresistant squamous cell carcinoma (SCC) cell lines. The 96-well plate clonogenic assay was used. Survival data were fitted to the linear quadratic model. The area under the curve (AUC), equivalent to mean inactivation dose (D ), was obtained with numerical integration. AUC ratios between single-dose radiation and fractionated radiation with or without paclitaxel were used to determine the SLDR of the cell lines and the effect of paclitaxel on it. Seven currently tested vulvar SCC cell lines were found to have a limited capacity of repairing sublethal damage (SLD). Only 3 of them presented SLDR of significance. The effect of concurrent radiation and paclitaxel was clearly additive when the radiation dose was fractionated in most of the cell lines. In addition, 2 of the cell lines having SLDR exhibited a trend toward losing the repair capacity when paclitaxel was present during the irradiation. In addition, the survival curve of the UM-SCV-1A cell line gave the impression of a true paclitaxel effect on SLDR. Paclitaxel used concurrently with fractionated radiation showed effectiveness on vulvar carcinoma. The effect was at least additive and could even be expected to abrogate the SLDR during split-dose radiation.

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“…The assay has been described earlier in detail (Grènman et al, 1989;Rantanen et al, 1994). The cells were harvested with trypsin-EDTA to obtain a single-cell suspension, counted and diluted in Ham's F-12 medium containing 15% FBS.…”

Section: Clonogenic Assaymentioning

confidence: 99%

Additive and supra-additive cytotoxicity of cisplatin-taxane combinations in ovarian carcinoma cell lines

et al. 1998

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SummaryThe purpose of this study was to compare the growth-inhibitory effect of cisplatin-paclitaxel with that obtained with a cisplatin-docetaxel combination and to assess the type of interaction. Concomitant use of taxanes and cisplatin was studied in seven human ovarian carcinoma cell lines, using the 96-well plate clonogenic assay. Chemosensitivity was expressed in terms of IC 50 values, the drug concentration causing 50% inhibition of clonogenic survival. The type of interaction was studied using the area under the survival curve ratios (AUC ratios) obtained by numerical integration. Comparison of the AUC ratio and the surviving fraction (SF) value after taxane alone was made using Student's t-test. The influence of the drug concentration was tested by one-way analysis of variance (Anova). A supra-additive or additive effect was seen when seven ovarian carcinoma cell lines were exposed to paclitaxel or docetaxel concomitantly with cisplatin. A supra-additive effect was found in four cell lines (UT-OC-3, UT-OC-4, UT-OC-5 and SK-OV-3) after simultaneous use of cisplatin with all docetaxel concentrations tested, and in two cell lines (UT-OC-4 and SK-OV-3) when cisplatin was used concomitantly with paclitaxel. A more pronounced supra-additive effect was seen with the combination of cisplatin and docetaxel. The degree of supra-additivity was dose dependent, with increasing synergy after a higher taxane dose. The data obtained in this study suggest that a supra-additive or additive effect can be achieved in ovarian carcinoma with the concomitant use of cisplatin and a taxane. for cervical cancer 30 and 5 years before the diagnosis of ovarian carcinoma. The donors of the UT-OC-1 and UT-OC-3 cell lines had not received any cytotoxic therapy before the establishment of the cell lines. Keywords Cell cultureBefore the experiments, the cells were kept in logarithmic growth in T25 culture flasks by passing weekly in Dulbecco's modified Eagle's minimal essential medium (DMEM) containing 2 mM L-glutamine, 1% non-essential amino acids, 100 U ml -1 streptomycin, 100 U ml -1 penicillin and 10% fetal bovine serum (FBS). Cells in mid-logarithmic growth (40-60% confluence) were used for the experiments and fed with fresh medium on the day before plating. Drug preparationCisplatin (Platinol) 0.5 mg ml -1 was diluted with growth medium to get a stock solution of 100 µg ml -1 . Final cisplatin dilutions of 0.05-0.6 µg ml -1 were used, and new stock solutions were made for each experiment. Paclitaxel (Taxol, kindly provided by BristolMyers Squibb) was initially dissolved in 0.9% sodium chloride to get a solution of 0.1 mM. Stock solutions were prepared in Ham's F-12 medium containing 10% FBS to obtain a solution of 100 nM, and stored at -40°C. Final dilutions of 0.4-5 nM paclitaxel were used for the experiments. Docetaxel (Taxotere, 807.9 mg, kindly provided by Rhone-Poulenc Rorer) was diluted in 1 ml of ethanol to obtain a stock solution of 0.1 mM and stored at -40°C. These solutions were further diluted in sterile water to obtain...

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Clonogenic cell assay for anchorage‐dependent squamous carcinoma cell lines using limiting dilution

Cited by 77 publications

References 26 publications

Establishment and characterization of human osteosarcoma cell lines with different pulmonary metastatic potentials

Establishment and characterization of human osteosarcoma cell lines with different pulmonary metastatic potentials

Paclitaxel combined with fractionated radiation in vitro: A study with vulvar squamous cell carcinoma cell lines

Additive and supra-additive cytotoxicity of cisplatin-taxane combinations in ovarian carcinoma cell lines

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