Epi5™ Episomal iPSC Reprogramming Kit - FAQs

我们提供Invitrogen CytoTune-iPS 2.0仙台病毒重编程试剂盒（Invitrogen CytoTune-iPS 2.0 Sendai Reprogramming Kit，货号A16517，A16518），这是一套非整合型的系统，通过仙台病毒载体将体细胞重编程为诱导性多能干细胞（iPSCs）。Invitrogen CytoTune-iPS 2.0仙台病毒重编程试剂盒包含了三种用于递送和表达关键遗传因子的Invitrogen CytoTune 2.0重编程载体，这些因子的表达对于将体细胞重编程为iPSCs的操作是必需的。只需要一次应用这些载体即可成功的实现重编程。

此外，我们还提供附着体iPSC重编程载体 (货号A14703) 或 Epi5附着体iPSC重编程试剂盒 (货号 A15960)。其同样是非整合型系统能将体细胞重编程为诱导性多能干细胞（iPSCs）。本品是六种或五种载体的混合物，为在无饲养层环境下生成iPSC提供优化的体系，具有非病毒介导、无基因组整合的特点。这些附着体iPSC重编程载体最初由Junying Yu与James Thomson开发（http://www.ncbi.nlm.nih.gov/pubmed/19325077），并经过Cellular Dynamics International的进一步优化，能够胜任大量不同种类的体细胞的重编程操作。

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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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) 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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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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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.

Find additional tips, troubleshooting help, and resources within our Cell Culture Support Center.