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.
Service includes:
- Lentivirus vector(s) expressing SV40 large and small T antigens
- Transduction, 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
Standard service fee for following samples:
- Primary Fibroblasts (from healthy or disease donor)
- Smooth Muscle Cells
- MEF cells
Please ask about restrictions or possible experimental services for the following samples or others not listed:
- Leukemia cells
- B cells
- Lymphoblastoid cells
- Peripheral blood mononuclear cells (PBMC)
- T cells
- Macrophages
- Retinal ganglion cells
- Tumor cells
- Chondrocytes
- Hepatocytes
- Myoblasts
- Osteoblasts
- Primary cell from ovary cancer tissue
Currently we are not accepting following samples:
- Neurons
- Dendritic cells
NOTE:
Some primary cells may encounter what’s known as a “crisis stage” after over ~25-30 passages. Therefore, truly immortalized cell lines must be cultured long-term to pass the “crisis stage.” Our guarantee includes the positive transduction and expression of T antigen in target cells. If required, ASC will add the long-term culture to test beyond the “crisis stage” option for additional charge, which will be estimated based on the growth properties of the transduced cells; please contact us for an additional quote.