STEMCCA Constitutive Polycistronic (OKSM) Lentivirus Reprogramming Kit

Research Category
Stem Cell Research

Tested to confirm the generation of iPS cells from p3 mouse embryonic fibroblasts and p6 human foreskin fibroblasts. Other cell types have not been tested and thus similar results can not be guaranteed.

The STEMCCA Constitutive Polycistronic (OKSM) Lentivirus Kit contains high titer polycistronic (OKSM) lentivirus and Polybrene transfection reagent that have been validated for the generation of mouse and human induced pluripotent stem cells from mouse embryonic fibroblasts (MEFs) and human foreskin fibroblasts (HFFs), respectively. The STEMCCA vector is comprised of the transcription factors Oct-4, Klf4, SOX-2, and c-Myc (OKSM), separated by the self-cleaving 2A peptide and IRES sequences (Sommer CA, 2009). The use of a single polycistronic lentiviral vector significantly improves reprogramming efficiencies and reduces the number of viral integrations.

Important note: the STEMCCA lentivirus reprogramming kits are provided in two formats, dependent on whether you are working with a human or mouse system. Human reprogramming experiments require a higher multiplicity of infection (MOI) than mouse experiments. Therefore, the larger, three-vial kits are required for human reprogramming experiments. If you are reprogramming mouse embryo fibroblast cells, you have the option to use either the one- or three-vial kit, depending on how many wells you want to infect. Reprogramming other rodent somatic cells may require more than one vial of lentivirus to perform at optimal MOI.

1. EF1α-STEMCCA (OKSM) Lentivirus: (Part number CS204279) One (1) vial containing 15 µL of high titer lentivirus.

2. Polybrene Transfection Reagent: (Part number TR-1003-50UL) One (1) vial containing 50 µL of 10 mg/mL stock.

Lentivirus is stable for at least 6 months when stored at -80°C. After first thaw, place immediately on ice and store in working aliquots to avoid further freeze thaws. Avoid freeze thaws as this will result in a decrease in the virus titer. Polybrene transfection reagent is stable for at least 1 year when stored at -20°C.