Fibroblasts Immortalization Service
Simian virus 40 (SV40) T antigen has been shown to be the simplest and most reliable agent for the immortalization of many different cell types and the mechanism of SV40 T antigen in cell immortalization is relatively well understood. Recent studies have also shown that SV40 T antigen can induce telomerase activity in the infected cells. Here the human fibroblasts were immortalized by introducing SV40 T antigen.
- Lentivirus vector(s) expressing SV40 large and small T antigens
- colony selection (up to 10 passages)
- Characterization (transgene expression by RT-PCR)
- Culture, cryopreservation
Required Materials: Customer-desired cells, 2 vials of typical storage size, and indication of appropriate growth medium and conditions. If your cells require medium other than DMEM and RPMI, we will need 1-2 L of medium and any applicable growth factors from you for the project. Also, please provide coated 6-well plates and T25 flasks if the cells require specially treated culture vessels.
Deliverables: Applied StemCell, Inc. will provide 2 vials (~1 x 10^6 cells/vial) of immortalized cells.
Timeline: 2-5 months depending on immortalizing method, source cells, and species
1. Sample A, 2. Sample B, 3. Sample A-SV40, 4. Sample B SV-40, 5. SV40 plasmid (positive control) 6. Ladder
- Poli, M. C., Ebstein, F., Nicholas, S. K., de Guzman, M. M., Forbes, L. R., Chinn, I. K., ... & Coban-Akdemir, Z. H. (2018). Heterozygous Truncating Variants in POMP Escape Nonsense-Mediated Decay and Cause a Unique Immune Dysregulatory Syndrome. The American Journal of Human Genetics, 102, 1-17. https://doi.org/10.1016/j.ajhg.2018.04.010