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Invitrogen™ Epi5™ Episomal iPSC Reprogramming Kit
Description
Features of the Epi5™ Episomal iPSC Reprogramming Kit include:
Create Transgene and Virus-free iPSCs
Episomal vectors are a well-described system for producing transgene-free, virus-free iPSCs, providing a source of iPSCs for all stages of your pluripotent stem cell research. Other reprogramming methods, such as lentivirus, contain transgenes that can integrate into the host genome, potentially disrupting the genome or causing unpredictable results.
As oriP/EBNA1 vectors, these episomal vectors contain 5 reprogramming factors (Oct4, Sox2, Lin28, Klf4, and L-Myc) and replicate extra-chromosomally only once per cell cycle. At this replication rate, the episomes are lost at a rate of approximately 5% per cell generation. This system shows enhanced iPSC generation through p53 suppression, and the inclusion of L-Myc has been shown to be more potent and specific then c-Myc during human iPSC generation (1).
Easy-to-Use System
This 2-vial kit provides the optimal system for episomal vector reprogramming and eliminates the need for use of small molecules in the reprogramming medium. The episomal vectors are introduced into the cell by electroporation. We recommend use of the Neon™Transfection System for electroporation, allowing flexibility in the volume and thus number of cells needed for reprogramming.
Flexibility in Selection of Media Systems
The Epi5™ Episomal iPSC Reprogramming Kit can be used with multiple cell types and media systems. Protocols for fibroblasts and CD34+ blood cells are available in either feeder or feeder-free based reprogramming.
1. Okita K, Matsumara Y, Sato Y, et al. A more efficient method to generate integration free human iPS cells. Nature Methods 8, 409-412, 2011.
Specifications
Specifications
| Product Line | Epi5 |
| Quantity | 1 kit |
| Promoter | CAG |
| For Use With (Application) | Stem Cell Engineering & Reprogramming |
| For Use With (Equipment) | Electroporator, Neon Transfection System |
| Product Type | Episomal iPSC Reprogramming Kit |
| Content And Storage | 2 tubes. Store at -5 to -20°C. |
| Replication Origin | pUC Origin |
| Reporter Gene | None |
| Vector | EBNA OriP |
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Frequently Asked Questions (FAQs)
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.
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.
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.
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.
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.
Safety and Handling
For Research Use Only. Not for use in diagnostic procedures.
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