Can I use pen/strep or other antibiotics for episomal reprogramming of cells?
The use of antibiotics is not recommended during episomal reprogramming, especially Fungizone antimycotic. If antibiotics must be used during the reprogramming process, pen/strep could be used. For established iPSCs (after reprogramming is complete), either antibiotic/antimycotic or pen/strep should be fine.
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Can I reprogram mouse cells with the Epi5 Episomal iPSC Reprograming Kit (Cat. No. A15960)?
No. The Epi5 Episomal iPSC Reprogramming Kit (Cat. No. A15960) was designed to reprogram human cells, not mouse cells. We currently only have data with this kit on reprogramming human cells, which aligns with the use of EBNA/oriP episomal vectors mainly in primate cells. Although rodent cells lack the oriP replicon and are not permissive, some studies replaced or modified oriP with other sequences to mediate replication and some literature reported the use of unmodified EBNA/oriP in certain hamster and mouse lines.
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When generating iPSCs using the Epi5 Episomal iPSC Reprogramming Kit (Cat. No. A15960), how confluent should my cells be on the day of transfection?
When generating iPSCs using the Epi5 Episomal iPSC Reprogramming Kit (Cat. No. A15960) cells should be approximately 75-90% confluent on the day of transfection (Day 0). Since over-confluency results in decreased transfection efficiency, we recommend replating overly confluent cells to achieve 75-90% confluency on the day of transfection.
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How many reprogramming experiments can I perform with the Epi5 Episomal iPSC Reprogramming Kit (Cat. No. A15960)?
The Epi5 Episomal iPSC Reprogramming Kit (Cat. No. A15960) provides enough material to process 20 reprogramming experiments (each reprogramming experiment requires 1 × 105 cells and 1 µL each of the Epi5 Reprogramming Vectors and the Epi5 p53 & EBNA Vectors). Each vial of vectors in the kit contains 20 µL, and you will use 1 µL (from each vial) per experiment. Before use, briefly centrifuge the thawed vectors to collect them at the bottom of the tube.
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Which kits do you offer for stem cell reprogramming?
We offer the Invitrogen CytoTune-iPS 2.0 Sendai Reprogramming Kit (Cat. Nos. A16517, A16518) which is a non-integrating system that uses Sendai virus vectors to reprogram somatic cells into induced pluripotent stem cells (iPSCs). The Invitrogen CytoTune-iPS 2.0 Sendai Reprogramming Kit contains three Invitrogen CytoTune 2.0 reprogramming vectors that are used for delivering and expressing key genetic factors necessary for reprogramming somatic cells into iPSCs. Only one application of the vectors is required for successful reprogramming.
Additionally, we offer the Episomal iPSC Reprogramming Vectors (Cat. No A14703) or Epi5 Episomal iPSC Reprogramming Kit (Cat. No A15960). This is a non-integrating system that reprograms somatic cells into induced pluripotent stem cells (iPSCs). These two products contain a mixture of six or five vectors designed to provide the optimal system for generating transgene-free and virus-free iPSCs in a feeder-free environment. Originally developed by Junying Yu and James Thomson (http://www.ncbi.nlm.nih.gov/pubmed/19325077) and further optimized by Cellular Dynamics International, these Episomal iPSC Reprogramming Vectors have proven successful in reprogramming a number of different somatic cell types.
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