Characterization and phenotypic variation with passage number of cultured human endometrial adenocarcinoma cells

Kato, Sumie; Espinoza, Natalia; Lange, Soledad; Villalón, Manuel; Cuello, Mauricio; Owen, Gareth I.

doi:10.1016/j.tice.2007.09.007

Cited by 19 publications

(15 citation statements)

References 27 publications

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“…At the same time, there are no documented data available for passaging of cardiomyocytes. Kato et al (2008) published results for the cultured human endometrial cancer line (CHEC), showing that with successive passages, cell morphology changes, together with expression levels for proteins such as cytokeratin, alpha-actin, or vimentin. Publications for other cancer cell lines include studies on the epithelial colorectal adenocarcinoma line, Caco-2, in which the changes in cell morphology occurring in successive passages were described, and an attempt was made to describe standards for culturing these cells to achieve repeatable results (Jahn et al 2011; Natoli et al 2012).…”

Section: Discussionmentioning

confidence: 99%

The effect of a number of H9C2 rat cardiomyocytes passage on repeatability of cytotoxicity study results

et al. 2016

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The embryonic cardiomyocyte cell line H9C2 is commonly used in numerous in vitro studies, including cardiotoxicity analyses of new drugs. So far no results were published for studies on cell parameters variability during the cell line ageing process. For this reason the aim of the study was to evaluate the effect of a number of H9C2 rat embryonic cardiomyocytes passages on repeatability of study results for selected cytotoxicity parameters, with doxorubicin as a model toxic agent. The cultures were passaged twenty-five times. Cells from passage 1, 5, 10, 15, 20 and 25 were treated with doxorubicin for 24 h. Then drug cytotoxicity was evaluated with the MTT test and additionally the nuclear factor erythroid 2–related factor (Nrf2) gene expression was examined. The analysis of oxidative stress intensity and cell morphology was also assessed. The microscopic appearance of cells indicates that untreated cardiomyocytes morphology changes as well as sensitivity to toxic effects increases with the number of passages. Also an increase in oxidative stress in cells occurs with further passaging of cardiomyocytes. Statistical significance of differences in conducted tests results depended on doxorubicin concentration but in many cases the H9C2 line was found to be a reliable in vitro model only for the first five passages. For this reason it is important to take into consideration that further culturing of cardiomyocytes may not ensure repeatability of study results due to the culture ageing.

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Section: Discussionmentioning

confidence: 99%

The effect of a number of H9C2 rat cardiomyocytes passage on repeatability of cytotoxicity study results

et al. 2016

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“…Primary human foreskin epithelial cells cultured in plastic dishes were found to increase in stiffness with passaging, with cells being twofold to fourfold stiffer after eight passages than were cells passaged fewer than three times56. Similarly, human epithelial breast carcinoma (MCF7) cells were found to stiffen with increasing passage number when cultured on glass coverslips57 and endometrial adenocarcinoma cells cultured in plastic dishes expressed more α-actin as a function of passage number, as they moved towards a stromal phenotype58. In each of these cases, the cells were grown on substrates with markedly different mechanical properties from native tissue, and long-lived changes in cytoskeletal properties and cytoskeletal organization were observed.…”

Section: Sensing the Mechanical Microenvironmentmentioning

confidence: 99%

Cell mechanics and the cytoskeleton

Fletcher

Mullins

2010

Nature

2,389

2,006

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The ability of a eukaryotic cell to resist deformation, to transport intracellular cargo and to change shape during movement depends on the cytoskeleton, an interconnected network of filamentous polymers and regulatory proteins. Recent work has demonstrated that both internal and external physical forces can act through the cytoskeleton to affect local mechanical properties and cellular behaviour. Attention is now focused on how cytoskeletal networks generate, transmit and respond to mechanical signals over both short and long timescales. An important insight emerging from this work is that long-lived cytoskeletal structures may act as epigenetic determinants of cell shape, function and fate.In a 1960 lecture, cell and developmental biologist Paul A. Weiss encouraged his audience to think of the cell as an integrated whole "lest our necessary and highly successful preoccupation with cell fragments and fractions obscure the fact that the cell is not just an inert playground for a few almighty masterminding molecules, but is a system, a hierarchically ordered system, of mutually interdependent species of molecules, molecular groupings, and supramolecular entities; and that life, through cell life, depends on the order of their interactions" 1 . This statement may be more relevant today than it was 50 years ago. Despite tremendous progress, fundamental gaps remain between our understanding of individual molecules and our understanding of how these molecules function collectively to form living cells. The sequencing of genomes outpaces characterization of the cellular components they encode and far exceeds our ability to reassemble these components into the types of complex system that can provide mechanistic insight into cellular behaviour. An even more difficult task is to connect the behaviour of cells in culture with that of more complex living tissues and organisms.Ever since muscle fibres were first examined under rudimentary microscopes in the seventeenth century, researchers have been motivated to understand how the process of self-organization generates dynamic, robust and elaborate structures that organize and 'animate' cells. The biological importance of establishing order over diverse length scales and timescales, as well as the challenges of understanding how systems of self-organizing molecules carry out cellular functions, is perhaps best illustrated by studies of the cytoskeleton.

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“…Consistent with this view, TF was also found to be highly expressed in three out of six primary USPC cell lines available for this study, using qRT–PCR and flow cytometry. The TF expression has been previously reported in endometrial endometrioid adenocarcinoma cell lines (i.e., the most common and less aggressive variant of human endometrial cancer; Kato et al , 2005, 2008). However, to our knowledge, the overexpression of TF in primary chemotherapy-resistant uterine serous tumours has not been previously studied.…”

Section: Discussionmentioning

confidence: 88%

hI-con1, a factor VII-IgGFc chimeric protein targeting tissue factor for immunotherapy of uterine serous papillary carcinoma

Cocco

Richter

et al. 2010

Br J Cancer

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Background:Uterine serous papillary adenocarcinoma (USPC) is a highly aggressive variant of endometrial cancer. Human immuno-conjugate molecule (hI-con1) is an antibody-like molecule targeted against tissue factor (TF), composed of two human Factor VII (fVII) as the targeting domain, fused to human immunoglobulin (Ig) G1 Fc as an effector domain. We evaluated hI-con1 potential activity against primary chemotherapy-resistant USPC cell lines expressing different levels of TF.Methods:A total of 16 formalin-fixed, paraffin-embedded USPC samples were evaluated by immunohistochemistry (IHC) for TF expression. Six primary USPC cell lines, half of which overexpress the epidermal growth factor type II (HER2/neu) receptor at 3+ levels, were assessed by flow cytometry and real-time PCR for TF expression. Sensitivity to hI-con1-dependent cell-mediated cytotoxicity (IDCC) was evaluated in 5-hour-chromium release assays. Finally, to investigate the effect of interleukin-2 (IL-2) on IDCC, 5-h 51Cr assays were also conducted in the presence of low doses of IL-2 (i.e., 50–100 IU ml−1).Results:Cytoplasmic and/or membrane TF expression was observed in all 16 (100%) USPC samples tested by IHC, but not in normal endometrium. High expression of TF was found in 50% (three out of six) of the USPC cell lines tested by real-time PCR and flow cytometry when compared with normal endometrial cells (NECs; P<0.001). Uterine serous papillary adenocarcinoma cell lines overexpressing TF, regardless of their high or low HER2/neu expression, were highly sensitive to IDCC (mean killing±s.d., 65.6±3.7%, range 57.5–77.0%, P<0.001), although negligible cytotoxicity against USPC was seen in the absence of hI-con1 or in the presence of Rituximab control antibody. The addition of low doses of IL-2 further increased the cytotoxic effect induced by hI-con1 against chemotherapy-resistant USPC.Conclusion:hI-con1 induces strong cytotoxicity against primary chemotherapy-resistant USPC cell lines overexpressing TF. The hI-con1 may represent a novel therapeutic agent for the treatment of patients harbouring advanced, recurrent and/or metastatic USPC refractory to standard treatment modalities.

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Characterization and phenotypic variation with passage number of cultured human endometrial adenocarcinoma cells

Cited by 19 publications

References 27 publications

The effect of a number of H9C2 rat cardiomyocytes passage on repeatability of cytotoxicity study results

The effect of a number of H9C2 rat cardiomyocytes passage on repeatability of cytotoxicity study results

Cell mechanics and the cytoskeleton

hI-con1, a factor VII-IgGFc chimeric protein targeting tissue factor for immunotherapy of uterine serous papillary carcinoma

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