File Name: cancer cell culture methods and protocols .zip
Passaging refers to the diluting of cells that have reached high confluence to supplement cells with fresh medium to enable continuous culture propagation. For mammalian cells, passaging should be performed when cells are toward the end of the logarithmic growth phase, so before they reach the stationary phase Figure 3. Passaging of cells at the stationary phase is not recommended because they tend to take longer to begin the logarithmic growth phase upon seeding. Additionally, the build-up of lactic acid in dense cultures may impact cell metabolism. Add detaching agent e. Resuspend cells in fresh medium, pipette thoroughly to obtain single cell suspension.
Metrics details. The use of patient-derived primary cell cultures in cancer preclinical assays has increased in recent years. The management of resected tumor tissue remains complex and a number of parameters must be respected to obtain complete sample digestion and optimal vitality yield. We provide an overview of the benefits of correct primary cell culture management using different preclinical methodologies, and describe the pros and cons of this model with respect to other kinds of samples. One important advantage is that the heterogeneity of the cell populations composing a primary culture partially reproduces the tumor microenvironment and crosstalk between malignant and healthy cells, neither of which is possible with cell lines. Moreover, the use of patient-derived specimens in innovative preclinical technologies, such as 3D systems or bioreactors, represents an important opportunity to improve the translational value of the results obtained.
Overview DOI: By mimicking crucial features of in vivo environment, including cell—cell and cell—extracellular matrix interactions, 3D cell culture enables proper structural. By mimicking crucial features of in vivo environment, including cell—cell and cell—extracellular matrix interactions, 3D cell culture enables proper structural architecture and differentiated function of normal tissues or tumors in vitro. Thereby 3D cell culture realistically models in vivo tissue conditions and processes, and provides in vivo like responses. Since its early days in the s, 3D cell culture has revealed important insights into mechanisms of tissue homeostasis and cancer, and accelerated translational research in cancer biology and tissue engineering. Antibody Data Search Beta. Authors: Zuzana Koledova 1.
Click on the following links to view the sample 1 protocol Part 2 Only and reagents list respectively, for conducting a Digesting DNA Reaction. When you have finished viewing the protocol, complete the theoretical lab assessment problem set. Click on the following links to see sample protocols for conducting a restriction enzyme digestion reaction and agarose gel electrophoresis. Protocol Samples Click on the following links to view the sample 1 Introduction, on modern techniques used to detect the RNA and protein output of a cell. You will focus on the RNA information for Hybridization and Northern Blotting as it applies to this subset, however, you will revisit the text in the Proteins Techniques module to better understand how one can conduct enzymatic assays to analyze gene expression by measuring the protein product of a gene.
It seems that you're in Germany. We have a dedicated site for Germany. The growing of cancer cells in the laboratory has been a powerful tool for studying and understanding the biology of cancer and the effects of drugs on cancer cells. In Cancer Cell Culture: Methods and Protocols, expert researchers describe in detail their most productive and up-to-date methods for growing cancer cells in the laboratory. Suitable for novice and experienced researchers alike, these readily reproducible techniques solve a diverse range of experimental problems. There are methods to characterize and authenticate cell lines, to isolate and develop specific types of cancer cells, and to develop new cell line models. Functional assays are provided to evaluate clonogenicity, cell proliferation, apoptosis, adhesion, migration, invasion, senescence, angiogenesis, and cell cycle parameters.
Unfortunately, due to limited availability of material and difficulties with protocols validation, primary models are rarely implemented into laboratory practice. We have compared protocols for primary cultures, differing in media components and plate coatings. Therefore, it may facilitate implementation of hardly available experimental models for a variety of analyses. Stable cancer cell lines have been considered valuable tools for analysis of cancer biology as well as applicable platforms for preclinical drug testing for many years [ 1 , 2 ]. Being relatively easy to culture basic media requirements, simple culturing protocols and having virtually limitless lifespan, these lines gained recognition among the majority of scientists. Since cancers are well known for high heterogeneity [ 3 ], stable cancer cell lines may be unable to adequately represent the complexity of these diseases [ 4 , 5 ].
Cancer Cell Culture: Methods and Protocols, Second Edition, Methods in Molecular Biology, vol. ,. DOI /_1.
It seems that you're in Germany. We have a dedicated site for Germany. With many recent advances, cancer cell culture research is more important than ever before. This timely edition of Cancer Cell Culture: Methods and Protocols covers the basic concepts of cancer cell biology and culture while expanding upon the recent shift in cell culture methods from the generation of new cell lines to the use of primary cells. There are methods to characterize and authenticate cell lines, to isolate and develop specific types of cancer cells, and to develop new cell line models.
Protocol 2 — Resuscitation of Frozen Cell Lines. Protocol 3 — Subculture of Adherent Cell Lines. Protocol 5 — Subculture of Suspension Cell Lines. Protocol 7 — Cryopreservation of Cell Lines. Protocol 8 — Testing for Bacteria and Fungi.
Request PDF | Cancer Cell Culture: Methods and Protocols | With many recent advances, cancer cell culture research is more important than ever before.
Human cancer cell lines are an integral part of drug discovery practices. However, modeling the complexity of cancer utilizing these cell lines on standard plastic substrata, does not accurately represent the tumor microenvironment. Research into developing advanced tumor cell culture models in a three-dimensional 3D architecture that more prescisely characterizes the disease state have been undertaken by a number of laboratories around the world. These 3D cell culture models are particularly beneficial for investigating mechanistic processes and drug resistance in tumor cells. Recent studies have demonstrated the potential of utilizing 3D cell culture models in drug discovery programs; however, it is evident that further research is required for the development of more complex models that incorporate the majority of the cellular and physical properties of a tumor.
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