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What Is an Immortalized Cell Line?
Immortalized cell lines are able to expand indefinitely while maintaining a reduced risk of death caused by mutations or artificial processes. The development of immortalized cell lines has facilitated stable materials for biomedical research, high-throughput screening, and lowered costs. It is important to note that immortalized cell lines are not equivalent to stem cells because stem cells have the potential to differentiate into multiple somatic cells, while immortalized cells tend to be terminally differentiated.
Creative Biolabs, as a biotechnology company, has devoted much effort to provide immortalized cell line services in various aspects. Through cutting-edge technologies and customized cell models, Creative Biolabs can provide you with a one-stop solution from biological mechanism to preclinical drug development, thus achieving higher quality, reproducible and efficient research results.
Definition of Immortalized Cell Lines
An immortalized cell line is a cell population that generally does not have the capacity to proliferate indefinitely, but due to mutations has acquired the ability to bypass normal cellular senescence and can proliferate indefinitely in vitro. Immortalized cell lines provide a continuous supply of cells and are commonly created by genetic mutation, carcinogenesis, or artificial methods (introducing immortalization genes). One of the first immortalized cell lines to be cloned was HeLa, which is derived from cervical cancer tissue of a patient.
Table 1 Characteristics of immortalized cell lines
| Characteristics | Scientific Mechanism | Research Significance |
|---|---|---|
| Unlimited Proliferation | Telomerase activity maintains telomere length, avoiding replicative senescence | Provides a long-term stable cell resource |
| Cloning Ability | Single-cell expansion forms a genetically uniform population | Ensuring experimental reproducibility |
| Phenotypic changes | Chromosomal abnormalities; metabolic pathway remodeling | Requires assessment of deviation from primary cells |
| Cultivation convenience | Adaptable to standard culture media, no complex growth factors required | Reducing experimental costs |
Technical Methods for Artificially Inducing Immortalized Cell Lines
The technical methods of artificially induced immortalization mainly include the expression of SV40 T antigen and telomerase (hTERT). The two methods promote the unlimited proliferation of cells by means of cell cycle regulation or telomerase activity activation, and make cells avoid senescent death.
SV40 T antigen is one of the most commonly used methods for cellular immortalization. It can bind to the p53 protein and inactivate it, so as to avoid cell division arrest and make cells enter the immortalized state. At the same time, the SV40 T antigen also has the function of activating telomerase activity and promoting cellular immortalization. For instance, the single use of SV40 T antigen or hTERT cannot immortalize colon stem cells, while the simultaneous use of them can greatly improve the clonal growth ability of colon stem cells.
Table 2 Viral gene transfection techniques and examples
| Method | Mechanism of Action | Application Examples |
|---|---|---|
| SV40 T Antigen | Inhibition of the p53/Rb tumor suppressor pathway | HEK293T |
| EB Virus | Activation of immortalization genes such as BCRF1 | Immortalization of B lymphocytes |
| HPV E6/E7 | Degradation of p53 protein | immortalization of cervical epithelial cells |
Telomerase activation is another important immortalization method. The expression of telomerase catalytic subunit hTERT can maintain telomere length, block telomere shortening at the end of chromosomes and avoid cell senescence. Research has found that cells immortalized by hTERT usually maintain normal phenotypes and stable karyotypes, and do not lead to tumor transformation. For instance, hTERT can be introduced into normal human cells, which can significantly prolong the life span of cells, and cells similar to primary cells in morphology and function can be obtained.
Table 3 Telomerase activation techniques and examples
| Method | Mechanism of Action | Application Examples |
|---|---|---|
| hTERT expression | reconstruction of the telomere maintenance system | human fibroblasts |
| Combined strategy | hTERT + SV40 T co-transfection | Human mesenchymal stem cells |
Figure 1 A schematic diagram of the telomerase remedial procedures.1,2
Representative Immortalized Cell Lines
Table 4 Representative immortalized cell lines and their applications
| Species | Cell line | Tissue origin | Application area |
|---|---|---|---|
| Human | HeLa | Cervical cancer | Virology/drug screening |
| HEK293 | Embryonic kidney | Gene therapy vector production | |
| Jurkat | T lymphocytes | Immunology research | |
| Mouse | 3T3 | Fibroblasts | Carcinogenicity testing |
| N2A | Neuroblastoma | Neural differentiation | |
| Monkey | Vero | Renal epithelium | Vaccine production |
Applications Of Immortalized Cells in Neuroscience
Models and Treatments for Neurodegenerative Diseases
Immortalized neural progenitor cells (e.g. HeLa cell line) can differentiate into neurons and glial cells and synthesize neurotrophic factors, which can be used as a potential tool for the treatment of neurodegenerative diseases such as Alzheimer's disease and Parkinson's disease. For instance, immortalized neural progenitor cells can be transplanted into the central nervous system (CNS) to repair damaged CNS and simulate the pathological process of the disease.
Immortalized astrocytes can be used to study the long-term recovery mechanisms after brain injury as they are the main regulators of neuronal nutrition support and synaptic activity.
Neural Regeneration and Stem Cell Research
Immortalized neural stem cells (e.g. tSA58, V-myc immortalized cell lines) have self-renewal ability and multipotent differentiation potential, which can be used for neural regeneration research. For example, RN33B cell lines can differentiate into neurons after transplantation, while tSA58 cell lines can differentiate into glial cells more than neurons. The research direction can be adjusted based on the differentiation results for specific treatment for different diseases.
Immortalized neural cell lines (e.g. 3T3, HEK293T) can stably express therapeutic genes through genetic engineering, which can be used for research on the growth and axon selection in the process of neuronal differentiation.
Drug Screening and Toxicity Testing
Immortalized cell lines (e.g. COS-1, CHO cells) can be used to produce expression systems for large-scale drug screening, such as generating erythropoietin for the treatment of anemia. Immortalized cells have advantages of rapid proliferation and stable phenotypes in high-throughput screening and thus can be used to test drug toxicity.
Immortalized neuronal cell lines (e.g. NSC-34, NSC-19) can be used to mimic the development of motor neurons, which can be used to study a model of chronic neuropathic pain.
Neural Development and Differentiation Mechanism Research
Immortalized cell lines (e.g. retrovirus-mediated Myc, SV40 T antigen-transfected cells) can be used to generate pluripotent neuronal cell lines for the study of neural progenitor cell differentiation regulation mechanisms. For example, retrovirus-mediated avian Myc gene expression can drive neural progenitor cells to co-express neuronal cells and glial cells at the same time.
Immortalized neural cell lines (e.g. N2A) can be used for the study of neuronal electrophysiological properties and neurotransmitter synthesis.
Disease Models and Personalized Medicine
Immortalized cell lines (such as immortalized human neural cells) can be used to establish patient-specific models for personalized medicine. For example, the genotype and phenotype of immortalized cells are almost the same as the primary cells, which can ensure the reproducibility of the experimental results.
Immortalized neural cell lines (such as mHypoA-Ast1) can be used to study the complex interactions of hypothalamic cells and the dysregulation in disease states.
Creative Biolabs is a specialist in the development of a range of neuro-based immortalized cell lines, including neurons, astrocytes, microglia and oligodendrocytes, and has extensive experience in the establishment and modification of these cell lines. We provide high-quality cell lines with validated identity, purity and long-term stability.
We offer cell line expansion, cryopreservation and custom cell line modifications with specified genetic backgrounds and gene editing capabilities. By using our state-of-the-art STEMOD™ technology platform, we provide comprehensive services for neuroscience research and CNS drug discovery.
Partner with Creative Biolabs to provide customized cell lines and world-class support to speed up your neuroscience project. Contact us today!
References
- Taheri, Mohammad, et al. "Hormonal Regulation of Telomerase Activity and hTERT Expression in Steroid-Regulated Tissues and Cancer." Cancer Cell International, vol. 22, no. 1, Aug. 2022, p. 258. DOI.org, https://doi.org/10.1186/s12935-022-02678-9.
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Created June 2025
For Research Use Only. Not For Clinical Use.