Culture of low passage colorectal cancer cells and demonstration of variation in selected tumour marker expression

Arul, Melanie; Roslani, April Camilla; Ng, Colin Leong Liong; Cheah, Swee Hung

doi:10.1007/s10616-013-9600-4

Cited by 5 publications

(4 citation statements)

References 34 publications

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“…This provides ease and speed of inventions. At the same time established cell lines represent a simplification of natural phenomena, as they are deprived of multilateral relations between different cell populations, microenvironment and responses of the host [7]. This can be partially overcome by culturing cells in complex three-dimensional systems or co-cultures, whaich better mimics natural surroundings in the host, retaining the ease of work and controlled conditions [8, 9].…”

Section: Introductionmentioning

confidence: 99%

Choosing the right cell line for renal cell cancer research

Brodaczewska¹,

Szczylik²,

et al. 2016

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Cell lines are still a tool of choice for many fields of biomedical research, including oncology. Although cancer is a very complex disease, many discoveries have been made using monocultures of established cell lines. Therefore, the proper use of in vitro models is crucial to enhance our understanding of cancer. Therapeutics against renal cell cancer (RCC) are also screened with the use of cell lines. Multiple RCC in vitro cultures are available, allowing in vivo heterogeneity in the laboratory, but at the same time, these can be a source of errors. In this review, we tried to sum up the data on the RCC cell lines used currently. An increasing amount of data on RCC shed new light on the molecular background of the disease; however, it revealed how much still needs to be done. As new types of RCC are being distinguished, novel cell lines and the re-exploration of old ones seems to be indispensable to create effective in vitro tools for drug screening and more.Electronic supplementary materialThe online version of this article (doi:10.1186/s12943-016-0565-8) contains supplementary material, which is available to authorized users.

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Section: Introductionmentioning

confidence: 99%

Choosing the right cell line for renal cell cancer research

Brodaczewska¹,

Szczylik²,

et al. 2016

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“…2 and 3). (2020) 10:9988 | https://doi.org/10.1038/s41598-020-66879-w www.nature.com/scientificreports www.nature.com/scientificreports/ 10-20, depending on cell line, as low passage reflects the characteristics of the primary tumor more closely and the malignancy kept 41,42 . Such experiments would be shorter than the one performed in this study, but sufficient enough to see, if cancer cell behavior is different form non-cancer cell.…”

Section: Discussionmentioning

confidence: 99%

Development of adaptive resistance to electric pulsed field treatment in CHO cell line in vitro

Polajžer

Miklavčič

2020

Sci Rep

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Pulsed electric field treatment has increased over the last few decades with successful translation from in vitro studies into different medical treatments like electrochemotherapy, irreversible electroporation for tumor and cardiac tissue ablation and gene electrotransfer for gene therapy and DNA vaccination. Pulsed electric field treatments are efficient but localized often requiring repeated applications to obtain results due to partial response and recurrence of disease. While these treatment times are several orders of magnitude lower than conventional biochemical treatment, it has been recently suggested that cells may become resistant to electroporation in repetitive treatments. In our study, we evaluate this possibility of developing adaptive resistance in cells exposed to pulsed electric field treatment over successive lifetimes. Mammalian cells were exposed to electroporation pulses for 30 generations. Every 5 th generation was analyzed by determining permeabilization and survival curve. No statistical difference between cells in control and cells exposed to pulsed electric field treatment was observed. We offer evidence that electroporation does not affect cells in a way that they would become less susceptible to pulsed electric field treatment. Our findings indicate pulsed electric field treatment can be used in repeated treatments with each treatment having equal efficiency to the initial treatment. Electroporation or pulsed electric field (PEF) treatment can cause increase in membrane permeability and allows molecules, for which the membrane is mostly impermeable, to cross. Two distinct outcomes of electroporation can be observed and used: reversible and irreversible electroporation. In reversible electroporation cells manage to repair the damage caused by electric pulses, i.e. cells survive. This has been widely used in different scientific fields for inserting molecules of interest into the cell. One of the more widely used applications is gene electro-transfer, which allows genetic manipulation. From a simple increased transfection this was developed into gene therapy 1,2 and DNA vaccination 3,4. While with gene therapy new genes and products are expressed within the host cell, DNA vaccination induces immune response of host organism. Combination of DNA vaccine injection and electroporation is most potent DNA delivery for subsequent immune response 5. Increased cell membrane permeability is also being exploited in electrochemotherapy (ECT). ECT is a combination of electric pulses and chemotherapeutic drug. Due to increased cell membrane permeability chemotherapeutic drug enter the cell which potentiates the cytotoxicity of the drug 6,7. ECT has been proved as a safe and efficient procedure for skin malignancies and now studies have been initiated to treat deep-seated tumors 8,9. The other possible outcome of electroporation is irreversible electroporation (IRE), where the damage to the cells is too extensive and leads to cell death. The use of irreversible electroporation as tissue ablation techn...

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“…Compared with previously reported protocols, additional antibiotics and washing steps were added as a precaution to prevent bacterial and mycoplasma growth as primary cultures from human tissues can suffer from contamination issues (Ebert et al, ; Shin et al, ; Vierck et al, ; Park et al, ; Perego et al, ; Vesey et al, ; Valente et al, ; Arul et al, ). Using this protocol, coating of culture flasks is not required and washed tumour/normal kidney tissue can be left overnight in culture medium at 4°C before further tissue processing with good success rate.…”

Section: Discussionmentioning

confidence: 99%

Establishment of epithelial and fibroblast cell cultures and cell lines from primary renal cancer nephrectomies

Yap

Ong

Morais

et al. 2019

Cell Biology International

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Renal cell carcinoma (RCC) is one of the most lethal urogenital cancers and effective treatment of metastatic RCC remains an elusive target. Cell lines enable the in vitro investigation of molecular and genetic changes leading to renal carcinogenesis and are important for evaluating cellular drug response or toxicity. This study details a fast and easy protocol of establishing epithelial and fibroblast cell cultures or cell lines concurrently from renal cancer nephrectomy tissue. The protocol involves mechanical disaggregation, collagenase digestion and cell sieving for establishing epithelial cells while fibroblast cells were grown from explants. This protocol has been modified from previous published reports with additional antibiotics and washing steps added to eliminate microbial contamination from the surgical source. Cell characterisation was carried out using immunofluorescence and quantitative polymerase chain reaction. Eleven stable epithelial renal tumour cell lines of various subtypes, including rare subtypes, were established with a spontaneous immortalisation rate of 21.6% using this protocol. Eight fibroblast cell cultures grew successfully but did not achieve spontaneous immortalisation. Cells of epithelial origin expressed higher expressions of epithelial markers such as pan‐cytokeratin, cytokeratin 8 and E‐cadherin whereas fibroblast cells expressed high α‐smooth muscle actin. Further mutational analysis is needed to evaluate the genetic or molecular characteristics of the cell lines.

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Culture of low passage colorectal cancer cells and demonstration of variation in selected tumour marker expression

Cited by 5 publications

References 34 publications

Choosing the right cell line for renal cell cancer research

Choosing the right cell line for renal cell cancer research

Development of adaptive resistance to electric pulsed field treatment in CHO cell line in vitro

Establishment of epithelial and fibroblast cell cultures and cell lines from primary renal cancer nephrectomies

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