Normal cells will die after a few rounds of proliferation because of cellular senescence. There are a few methods to turn a primary cell to immortal so that the cells can undergo infinite cell divisions or large rounds of doubling in culture medium.
- One Cell Immortalization method is to repress the genes that control cell cycle such as such as retinoblastoma (Rb) and p53 genes (the tumor suppressor proteins). Many virus induced cancer cells have their viral genes repress “tumor suppressor proteins” and became immortal. Over-expression of such viral genes can turn primary cells into immortal. The most common of these is the Large T-antigen of the simian virus (SV-40). The others of these are: Human Papilloma Virus (HPV)’s E6 and E7 genes, Epstein-Barr Virus (EBV), E1A gene of human adenovirus type 5, Ras_V12 mutant, and more. Also, siRNAs that knockdown p53 or Rb can also be used for this route.
- Another Cell Immortalization method is to over-express the genes that confer immortality. The most well-known gene is Telomerase (hTERT) which is often found over-expressed in human tumors. Other those genes are HOX genes, CDK4 and more.
Depending on your cell types, you can combine both methods above to increase the immortalization efficiency or to immortalize a larger number of cells. Please be advised, after the immortalization, the phenotype of your primary cell may or may not be changed. And the immortalization is not necessarily oncogenesis. But, there are difference between primary cells and immortalized cells in terms of experimental model.
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