You are here: Home > Products > Lentiviral system >

Cell Immortalization Kits

Primary cells undergo finite number of population doublings, and reach a state called replicative senescence after limited cell divisions. GeneCopoeia’s Cell Immortalization Kits use recombinant lentivirus carrying genes, such as the simian virus 40 (SV40) T antigen, and the human telomerase reverse transcriptase protein (hTERT) to induce immortalization and significantly prolong primary cells lifespan.

 

Buy Catalog# Promoter Selection Description Price
LPP-Sv40T-Lv105-100-C CMV Puromycin SV40 large T antigen Purified Lentifect™ Lentiviral Particles (25 µl x 4 vial) $725
LPP-Q0450-Lv105-100-C CMV Puromycin hTERT Purified Lentifect™ Lentiviral Particles with Puromycin marker (25 µl x 4 vial) $725
LPP-T1758-Lv105-100-C CMV Puromycin CDK4 Purified Lentifect™ Lentiviral Particles with Puromycin marker (25 µl x 4 vial) $725
LPP-T1371-Lv105-100-C CMV Puromycin HOXB8 Purified Lentifect™ Lentiviral Particles with Puromycin marker (25 µl x 4 vial) $725
LPP-P0078-Lv105-100-C CMV Puromycin HOXA9 Purified Lentifect™ Lentiviral Particles with Puromycin marker (25 µl x 4 vial) $725
LPP-Z5789-Lv105-100-C CMV Puromycin HOXA10 Purified Lentifect™ Lentiviral Particles with Puromycin marker (25 µl x 4 vial)

$725

LPP-CMYC-Lv105-100-C CMV Puromycin c-Myc Purified Lentifect™ Lentiviral Particles with Puromycin marker (25 µl x 4 vial) $725

 

 

GeneCopoeia Lentifect™ Lentiviral Particles are produced from a standardized protocol using purified plasmid DNA and the proprietary reagents, EndoFectin™ Lenti (for transfection) and TiterBoost™ solution. The protocol uses a third generation self-inactivating packaging system meeting BioSafety Level 2 requirements.

The Lentifect™ particles include a CMV promoter for efficient expression of the immortalization inducer gene in target cells and use a puromycin resistance marker for selection of stably transduced cells.

Provided as 4 vials of 25 µl of lentiviral particles with titers of ~1 x 108 TU/ml. Lentifect™ particles are shipped on dry ice and must be stored at –80°C immediately upon receipt. Avoid repeated freeze-thaw cycles as this will reduce titers.

The lentiviral expression construct was validated by full-length sequencing, restriction enzyme digestion and PCR-size validation using gene-specific and vector-specific primers. Product is confirmed free of bacteria, fungi and common Mycoplasma contamination.

 

Cell Immortalization Protocol

  • Plate target primary cells in a 6-well plate at density of 0.5-2 x105 cells per well
  • Thaw the lentiviral particles in 37 ºC water bath and immediately remove it from the water bath once thawed
  • Transduce the cells in a 6-well plate with immortalizing lentivirus or GFP-lentivirus (positive control) at a MOI of 10-100. Dilute the lentivirus with fresh complete medium if cytotoxicity is a problem
  • The next day, remove viral supernatant and add appropriate complete growth medium to the cells and incubate at 37 ºC
  • After 72 hours incubation, passage the transduced cells in complete medium supplemented with the appropriate amount of selection antibiotic
  • Pick antibiotic-resistant colonies and cultivate continuously in complete medium with antibiotics to test the immortalization
  •