SV40 large T antigen inactivates both p53 and pRb tumour suppressor pathways. This strongly drives cell proliferation, but introduces oncogenic perturbation. hTERT overexpression extends replicative lifespan by maintaining telomere length, without directly inactivating p53 or pRb. This makes it a lower-perturbation strategy that better preserves normal cell physiology for many applications.
Cells & Cell Culture
Cell Immortalisation Kits and Reagents
Cell immortalisation kits and reagents for renewable, phenotype-preserving cell stocks
Immortalisation converts a primary cell population into a stable, expandable line that keeps dividing well beyond the natural Hayflick limit. The genetic strategy you choose determines how closely your resulting cells still behave like the original tissue.
The two most-used genetic triggers are SV40 Large T antigen and hTERT. SV40 Large T inactivates p53 and pRb to override senescence checkpoints. hTERT extends telomere length without suppressing tumour suppressors. This can better preserve your differentiated phenotype in epithelial and stromal cells. EBV-based transformation is a third route, used for example in B-lymphocytes.
Lentiviral constructs let you deliver immortalising genes for stable genomic integration. Wild-type EBV virus reagents give you B-cell transformation instead.
CDK4 offers a further route you can pair with hTERT. The combination gives you a more authentic model than hTERT alone, and suits epithelial cells that resist standard hTERT immortalisation. HPV E6 and E7 proteins take a similar path to SV40: they inactivate p53 and Rb, and are widely used to immortalise keratinocytes and other epithelial cell types.
Choosing the right immortalisation approach
- Need a renewable supply of a specific primary cell type, already immortalised: you can check the catalogue of pre-made immortalised cell lines first. Hundreds of human, mouse, rat, and canine lines are available to you, including hepatocytes, endothelial cells, and neural subtypes.
- Immortalising epithelial or stromal cells where phenotype preservation matters: an hTERT-based vector is the lower-perturbation option for you. It extends proliferative capacity without fully inactivating p53 or pRb.
- Working with B-lymphocytes: EBV transformation is the classical route, and remains the most efficient method for this lineage.
- Explore every immortalisation route in more depth in the free cell immortalisation handbook from our partner abm. It covers SV40, hTERT, CDK4, HPV E6/E7, EBV, and more, with protocols you can follow.
Applications
Sustainable cell line generation
Immortalised lines give you a renewable, consistent supply of a defined cell type, replacing repeated donor-dependent isolations in pharmacology studies.
Tissue-specific disease modelling
Immortalised hepatocytes, endothelial, renal, and neural subtypes retain lineage-specific expression, helping you study drug metabolism and pathology.
B-lymphocyte immortalisation
EBV transformation of peripheral blood B-lymphocytes is the established route, giving you proliferating B-cell lines with antigen-specific immunoglobulin expression.
Stable transgene expression
Lentiviral delivery of an immortalising gene can carry a reporter cassette in the same construct, linking proliferation with stable transgene expression.
Reduced reliance on tissue collection
Once a line is established from a validated primary cell isolate, you avoid the variability and donor burden of sourcing fresh tissue each time.
Consistent source for screening
A genetically stable line gives reproducible results across compound panels, supporting cell-based assay workflows where variability would confound your results.
Frequently asked questions
This depends on the immortalisation method and cell type. hTERT-immortalised lines often retain differentiated functions closely matching their primary counterparts. SV40-immortalised lines may show altered signalling and gene expression. We recommend validating key functional endpoints against freshly isolated primary cells before using immortalised lines as primary cell surrogates.
Serial passaging beyond the normal Hayflick limit, with maintained growth rate and viability, confirms extended proliferative capacity. Transformation assays, such as anchorage-independent growth in soft agar or focus formation, assess whether immortalisation has produced full oncogenic transformation. This is relevant when characterising lines for use in cancer biology versus normal physiology studies.
EBV transformation efficiently immortalises mature naive and memory B-lymphocytes from peripheral blood, via the EBV latent infection programme. It is less efficient for plasmablasts and plasma cells. The resulting lymphoblastoid cell lines express a restricted set of latent EBV antigens. You should distinguish these from primary B-cells in functional immunological assays.
Product catalogs
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Tell us what you are looking for — antibodies, kits, proteins, or supplies — and our team will point you to the best catalog filters, suppliers, and product matches for your workflow.
