Squamous cell carcinomas of the head and neck cultured in floating collagen gels: 1. The maintenance of stromal and epithelial elements in vitro without fibroblast overgrowth
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The study of the molecular biology of head and neck squamous cell carcinomas has been heavily reliant on the analysis of cell lines. This is largely because the maintenance of primary cell cultures is difficult. However, being monoclonal, cell lines are not representative of the primary tumor because of the loss of tumor cell heterogeneity. We report a technique for primary culture of squamous cell carcinomas with maintenance of epithelial and stromal cell components without overgrowth of the fibroblast cells. Phenotypic markers for fibroblasts and squamous cells were present up to 45 days after initiation of culture, and expression of epidermal growth factor receptor and involucrin in cultures paralleled that in the primary tumor. In vivo, tumor stromal elements are thought to play an important role in the support of epithelial cell growth. In the collagen gel system the preservation of the stromal cell component likely improves culture viability and growth. More importantly, this culture system allows the in vitro tumor to more accurately reflect the tumor from which it was derived, and it permits the study of primary squamous cell carcinomas under in vitro conditions.Objective To observe the effects of IP6 on human pancreatic adenocarcinoma cell line PC-3 and human pancreatic primary culture cell proliferation and differentiation in vitro.Methods Direct counting cells for cell growth curve,MTT test,flow cytometry test cell apoptosis were used to observe cell proliferation and differentiation.Results IP6 inhibited the growth of human pancreatic adenocarcinoma cell line PC-3 in a time and dose dependent manner.The effect was biggest when the level was 10mmol/L,but little difference with 5mmol/L and 10mmol/L.When less than 5mmol/L,IP6 almost can not effect the growth of human pancreatic primary culture cell;only little inhibition in 5mmol/L,and it was obviously inhibited in 10mmol/L.Conclusion When 5mmol/L,IP6 is the best inhibited density,which could kill tumor cell utmost and interfere normal cell in the lowest level.
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Bromodeoxyuridine
Hep G2
Thymidine
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<p>Most representative microscop photograph of human fibroblast cell line. (E) Cell viability was performed by MTS Assay on CCD- 112Sk cell line, in order to evaluate if complex affects negatively healthy cells. We have not observed toxic effect(incubation time 96 hrs) on fibroblast cultures, neither with complex nor virus alone and Carnosine6K alone (virus:100 VP/cell, Carnosine6K: 8mM).</p>
Viability assay
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Abstract: Involucrin is a protein precursor of the epidermal cornified envelope. Although expression of the human protein has been documented extensively, studies in the mouse have been hampered by a shortage of good antibodies. We describe the production of recombinant mouse involucrin and preparation of rabbit antisera to the protein that work well by immunohistochemistry and Western blotting. We confirm that in normal mouse epidermis the onset of involucrin expression is in the upper spinous layers and inner root sheath of the hair follicle. Involucrin was also detected in the differentiating epithelial cells of normal tongue, oesophagus and bladder. Involucrin was expressed in a subpopulation of mouse keratinocytes cultured in standard or low calcium medium and the proportion of involucrin‐positive cells increased during suspension‐induced terminal differentiation. Western blotting of keratinocytes from several inbred mouse strains revealed a remarkable heterogeneity in the electrophoretic mobility of involucrin, reflecting inter‐strain variation in the number of tandem repeats in the protein. In the hyperproliferative epidermis of healing wounds involucrin was expressed in most of the suprabasal layers. In epidermal papillomas and carcinomas involucrin expression correlated well with degree of histological differentiation. The sites of expression of the mouse protein were thus the same as those previously reported for human involucrin. With the development of the new antibodies we anticipate that involucrin will become as widely used a marker of keratinocyte differentiation in the mouse as it is in the human.
Loricrin
Epidermis (zoology)
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Cell Proliferation Profile of Five Human Uveal Melanoma Cell Lines of Different Metastatic Potential
<i>Objective:</i> The aim of this study was to establish a proliferation profile of uveal melanoma cell lines, using different methods, and to compare it with their previously determined metastatic potential (MP). <i>Methods:</i> Four human uveal melanoma and one transformed human uveal melanocytic cell line were ranked according to proliferation profiles. The proliferation profiles of the cell lines were compared to their MPs, which were previously determined from an immunosuppressed rabbit model. <i>Results:</i> Ranking of the cell lines using pulse labeling with tritiated thymidine was similar to the MP of the cell lines. <i>Conclusion:</i> The correlation between the proliferative rate of the uveal melanoma cell lines and their previously determined MP resulted in the proposal of a new classification scheme: high proliferation/high MP, low proliferation/low MP, and high proliferation/no MP. High proliferative capacity of a cell line did not necessarily confer MP; therefore, further cellular functions/adaptations must be required for tumor cell dissemination, survival, and growth at a metastatic site.
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WI-38
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Abstract Previous combo treatment experiments on BLM human melanoma cell line identified two combos vit-A + RU-486 and Cur+P+RU as the best combinations to decrease melanoma cell growth as well as IL-8 secretion in -vitro. In order to confirm this finding, experiments were repeated on two other human melanoma cell lines 1205Lu and WM238. Initial single or solo compound treatment on 1205Lu cells resulted in cell growth in the range of 37–59% compared to untreated control 1205Lu cell growth at 100%. A double combo treatment with combinations such as D+P, D+RU, A+P, A+RU, A+D resulted in cell growth in the range of 17 to 37% on the 1205Lu cell line. Whereas on WM238 cell line resulted in cell growth in between 18–22%. A triple combo such as combinations of three compounds (Cur+P+RU, Cur+A+P, Cur+D+RU) resulted in cell growth in the range of 19-30% on 1205Lu cells, whereas on WM238 cells resulted in between 19–20% cell growth. Elisarray of the supernatants from the combos which showed a significant decrease in cell growth revealed the regulation only in IL-8 secretion. IL-8 secretion in the supernatants of combos treated with A+RU and Cur+P+RU was17.5 and 8. 0 pg/ml respectively on the 1205Lu cell line. Whereas, IL-8 secretion was 0.515 and 0.926 pg/ml respectively in the supernatants of A+RU and Cur+P+RU combo treatments on the WM238 cell line. Conclusion Based on cell growth, Elisarray and IL-8 quantitation, two combos A+RU and Cur+P+RU also fared well on the two other cell lines 1205Lu and WM238. Presentation: No date and time listed
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S ummary . We have demonstrated that normal human bone marrow stromal cells can be induced to produce high levels of β‐interferon (IFN). One of three randomly selected human stromal cell lines produced β‐IFN in similar amounts to one of the best β‐IFN producer cell lines (MG‐63 osteosarcoma cells). The marrow stromal cell lines did not produce γ ‐IFN, though a low level of α‐IFN was apparently produced. The stromal cell lines differ from usual producer cell lines in that they can be subcultured for a much longer growth period and also maintain the ability to producer IFN. The cells are karyotypically normal and are not virus transformed. Such cell lines may be useful in the production of human IFN as well as allowing studies on the role of IFN in stromal cell‐haemopoietic cell interactions.
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