Non-Integrating iPSC Generation
Allele Biotechnology's NextGen mRNA platform allows for easy, hightly efficient, feeder free and xeno free iPSC generation in less time than every before. We are offering three unique ways for you to harness this breakthrough in iPSC reprogramming:
- 6F mRNA reprogramming premix: A validated ready-to-use mRNA mixture with 6 factors including an engineered Oct4 for accelerated iPSC induction
- Custom iPSC Generation Service: A fully customized all encompassing service for your specific cell type, including target cell culture, reprogramming and validation
- IVT Templates: Validated ready-to-use IVT templates for generating your own mRNA, reprogramming and directed differentiation factors are available
- Warren, L. et al. Feeder-Free Derivation of Human Induced Pluripotent Stem Cells with Messenger RNA. Scientific Reports, srep00657 (2012)
Induced Pluripotent Stem Cells (iPSC): Stem Cell Generation
The direct reprogramming of somatic cells to pluripotency was first accomplished in 2006. Adult mouse fibroblasts were converted to iPS Cells (induced pluripotent stem cells) through ecotropic expression of a select group of transcription factors. In 2007, direct reprogramming was achieved in human cells. The generation of iPS from differentiated adult cells has vast therapeutic implications, particularly in the context of pharmaceutical screening and cellular replacement therapies. Extensive research has been conducted in search of efficient ways for stem cell generation.
- Takahashi, K., and Yamanaka, S. (2006). Induction of pluripotent stem cells from mouse embryonic and adult Fibroblast cultures by defi ned factors. Cell 126, 663–676.
- Takahashi, K., Tanabe, K., Ohnuki, M., Narita, M., Ichisaka, T., Tomoda, K., and Yamanaka, S. (2007). Induction of pluripotent stem cells from adult human fibroblasts by defined factors. Cell 131, 861–872.
- Maherali, N., and Hochedlinger, K. (2008). Guidelines and techniques for the generation of Induced pluripotent stem cells. Cell Stem Cell 3
Nanog, Oct4, and Rex1 promoters have been reported to display ES/iPS cell specific expression under undifferentiated states. Allele Biotech provides pre-packaged lentiviral fluorescent protein (FP) reporters driven by all three promoters to confirm the state of the reprogrammed iPSCs. A GFP or RFP gene was cloned behind these ES-specific promoters to offer convenient fluorescence tracking of undifferentiated cells.
We also provide the iPS Cell Identification CR Primer Kits. Instead of ordering individual oligos, complete sets of primers, as precisely defined and tested in Takahashi et al. 2006, are conveniently and sufficiently provided for 50 reactons for each gene.
For iPSCs identification antibodies, chech the Antibody group or search in products.
- Da Yong WU, Zhen YAO (2005). Isolation and characterization of the murine Nanog gene promoter. Cell Research, 15 (5): 317–324.
- Rachel Eiges, Maya Schuldiner..et.al (2001). Establishment of human embryonic stem cell-transfected clones carrying a marker for undifferentiated cell. Current Biology 11: 514–518.
- Guangjin Pan, Jun Li, Yali Zhou, Hui Zheng, and Duanqing pei (2006). A negative feedback loop of transcription factors that control stem cell pluripotency and self renewal. ASEB Journal 20: E1094~E1102
Note: The specificity of these promoters is based on scientific publications, which are subject to debate.
Human embryonic stem (ES) cells or induced pluripotent stem (iPS) cells promise to serve as an unlimited source for transplantation or tissue-specific differentiation. However, obtaining and maintaining stem cells can be difficult for multiple reasons. For instance, most stem cell lines tend to spontaneously differentiate in culture, and even if the cells form stem cell-like colonies, they may be of a heterogeneous population. There have been a number of publications using murine Oct-4, Nanog, and Rex-1 promoters driven fluorescent proteins as markers for pluripotency tests [1-3]. Allele Biotech provides, under its iPS product line, packaged and validated lentiviral particles that would insert these 3 promoter-FP reporters into the stem cells. Although currently these promoters are of mouse sequences, their use in human stem cells has been reported.
To learn more about pluripotency reporters, email us at firstname.lastname@example.org or check out the iPS Pluripotency Reporter blog entry.