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Tumor-Derived Cell Lines

Tumor-derived cell lines are cell lines that have been established from cancerous tissues and serve as invaluable models in cancer research. They offer excellent opportunities to study cancer biology, test drug efficacy, and develop personalized medical treatments.

Generation of Tumor-Derived Cell Lines

I. Isolation

Cell line generation starts with collecting a tumor sample through biopsy or surgical resection. At this part, it is important that ethical and regulatory approvals are in place. The tumor sample is then transported to the laboratory in a sterile medium, often supplemented with antibiotics to reduce the probability of contamination.

Once in the lab, the tumor tissue is carefully sliced into small fragments using sterile instruments. To dissociate the tissue into single cells, enzymatic digestion is performed using enzymes such as collagenase, trypsin, or dispase. This step is usually done along with mechanical agitation. The cell suspension obtained is then filtered using a cell strainer to remove debris. This is followed by washing with phosphate-buffered saline (PBS) or culture

II. Culturing

The cells are then seeded into a suitable culture medium designed to mimic the tumor microenvironment. This typically includes specialized formulations such as Dulbecco’s Modified Eagle Medium (DMEM) or Roswell Park Memorial Institute (RPMI)-1640, supplemented with fetal bovine serum (FBS), growth factors, and other additives that support tumor cell proliferation.

Cultures are maintained under optimal conditions, at a temperature of 37°C with 5% CO₂, while monitoring daily for attachment and growth. Tumor cells are generally heterogeneous, and selective pressures in culture favor the growth of adaptable cells. Subculturing is performed when the cells reach 70-80% confluence. Trypsin or similar agents to detach them for reseeding is often used during subculturing.

III. Passaging

In order to establish a stable cell line, it is necessary to performed repeated passaging. During this phase of cell line generation, it is critical to make sure at this part that contamination testing is done. Also, the integrity of the cell line should be ensured through characterization. Techniques such as immunohistochemistry, flow cytometry, and genetic profiling are used to confirm the tumor origin, subtype, and relevant biomarkers.

How to Generate a Stable Cell Line

A stable tumor cell line is necessary in biomedical research for obvious reasons but the two most important are:

  1. Consistency and Reproducibility: Stable cell lines provide a reliable and uniform source of biological material that can be used across multiple experiments. This consistency ensures that results are reproducible, which is crucial for validating findings in scientific research.
  2. Genetic Stability: A stable cell line maintains its genetic integrity over multiple passages, ensuring that experimental results accurately reflect the properties of the original tumor. This genetic fidelity allows for more reliable studies related to mutations and other genetic alterations associated with cancer.

Steps to Generate a Stable Tumor Cell Line

  1. Selection of Cell Line:
    • Commonly used cell lines include HEK293T, HeLa, and various cancer-specific lines. 
  2. Transfection or Transduction:
    • Transfection: Introduction of a plasmid containing the gene of interest along with a selectable marker into the cells.
    • Transduction: Use of viral vectors to deliver the gene of interest into the cells.
  3. Recovery and Selection: Cells are allowed to recover for 48-72 hours and then applied antibiotics to eliminate non-transfected cells.
  4. Isolation of Clonal Cell Lines: Surviving cells are subjected to single-cell isolation allowed to grow into clonal colonies.
  5. Characterization and Validation: Western blotting or quantitative PCR (qPCR) are used to confirm expression of the gene of interest to characterize stable cell line.
  6. Long-term Culture and Maintenance: The stable cell line is maintained under optimal culture conditions, ensuring proper passage and growth rates to prevent genetic drift or loss of expression over time (Tandon et al., 2018).

IV. Cryopreservation

The last step of the cell line generation process is cryopreservation, wherein cells are suspended in a cryoprotectant solution, such as 10% dimethyl sulfoxide (DMSO) in FBS. The sample is then stored in liquid nitrogen for long-term use (Geraghty et al., 2014).

Uses of Tumor-Derived Cell Lines

Cancer Biology Research

Tumor-derived cell lines are commonly used in cancer biology research to understand tumor mechanisms. Using these cell lines, scientists can study the basic characteristics of cancer cells such as cell differentiation, proliferation, metastasis and their interactions with the microenvironment.

Cancer biology research also involves genetic and epigenetic studies. This entails the use of tumor cell lines to investigate oncogene activation, tumor suppressor gene activation, and other genetic transformations that drive cancer behavior.

Drug Discovery and Development

Tumor-derived cell lines are also invaluable in drug discovery and development. Using tumor cell lines, thousands of drug candidates with potential anti-cancer effects can be rapidly tested through high-throughput screening. These cell lines are also used to investigate drug resistance mechanisms, allowing researchers to identify techniques to overcome or prevent resistance to drug and therapies.

Genetic Studies

Tumor-derived cell lines have been used in helping identify genetic mutations that are characteristic of specific kinds of cancers. Cell lines are also useful for gene editing to study gene function or develop models possessing defined genetic mutations using techniques such as CRISPR-Cas9.

Biomarker Discovery

Tumor cell lines also enable the identification of predictive biomarkers that predict cancer cells’ reaction to specific therapies. These cell lines can also help find prognostic markers that provide information on disease outcomes.

Immunotherapy Research

Tumor cell lines are the logical specimen when investigating how cancer cells evade immune detection. They are also key to understanding how immune cells can be activated to fight cancer. Tumor cell lines have also been indispensable during evaluation of the effectiveness of immune-based therapies such as CAR-T and checkpoint inhibitors.

Radiotherapy Research

Tumor cell lines enable researchers to observe how cancer cells respond to radiation and to find the factors that either enhance sensitivity or enhance resistance which are highly useful when developing radiotherapy regimes.

Cancer Modelling

Recently, 3D culture models, organoids and sphenoids have been using tumor cell lines to recreate the behavior and architecture of tumors in vivo. Tumor cell lines also provide information about the heterogeneity of tumors and mechanisms behind different cancer types. Have you decided on what type of tumor cells you need for any of the above studies?

Assays Used for
Studying Tumor-Derived Cell Lines

Proliferation Assays

Proliferation Assays (metabolic activity assays, DNA synthesis assays, ATP concentration assays, flow cytometry, etc.)

Migration and Invasion Assays

Drug Screening Assays

3D Culture Models

Reporter Gene Assays

RT-PCR and Western Blotting

Matched Samples

PBMCs

PBMCs (peripheral blood mononuclear cells) | Click here to read more about PBMCs.

PBMCs

PBMCs (peripheral blood mononuclear cells) | Click here to read more about PBMCs.

Biofluids

Biofluids (sputum, saliva, aspirate, swabs, urine, stool, synovial fluid, cerebrospinal fluid) | Click here to learn about our Animal Clinical Samples

Biofluids

Biofluids (sputum, saliva, aspirate, swabs, urine, stool, synovial fluid, cerebrospinal fluid) | Click here to learn about our Animal Clinical Samples

Normal Adjacent Tissue

Normal Adjacent Tissue

Lymph nodes

Lymph nodes

More on request

More on request

Available Services for
Characterization

FACS (fluorescence-activated cell sorting)

qPCR (quantitative Polymerase Chain Reaction)

RNAseq (RNA sequencing)

Sterility

HLA (human leukocyte antigen) typing

More on request

We are About Speed & Quality

Cells are isolated from sourced samples on demand.

Fresh samples are processed at our facilities and cryopreserved cells are shipped on dry ice or liquid nitrogen worldwide.

We typically ship cells that are in stock within two days.

We ship cells Europe-wide within 24-48 hours, and can ship intercontinentally with dry shipper.

How to Order

To place an order or inquire about our services, please contact our sales team through the contact form on our website or via email by clicking Contact Us below. We’ll work with you to better understand your specific needs and provide a customized solution.

References

Geraghty, R. J., Capes-Davis, A., Davis, J. M., Downward, J., Freshney, R. I., Knezevic, I., Lovell-Badge, R., Masters, J. R., Meredith, J., Stacey, G. N., Thraves, P., Vias, M., & Cancer Research UK (2014). Guidelines for the use of cell lines in biomedical research. British journal of cancer111(6), 1021–1046. https://doi.org/10.1038/bjc.2014.166

Tandon, N., Thakkar, K. N., LaGory, E. L., Liu, Y., & Giaccia, A. J. (2018). Generation of Stable Expression Mammalian Cell Lines Using Lentivirus. Bio-protocol8(21), e3073. https://doi.org/10.21769/BioProtoc.3073