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.

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

1. Sample A, 2. Sample B, 3. Sample A-SV40, 4. Sample B SV-40, 5. SV40 plasmid (positive control) 6. Ladder

 

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