Pluriton™ Reprogramming Medium
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The Stemgent Pluriton Reprogramming Medium is a defined, xeno-free medium optimized for mRNA based cellular reprogramming of human cells. Pluriton Reprogramming Medium has been functionally validated, using the Stemgent mRNA Reprogramming Factor Set: hOKSML, to support the generation of induced pluripotent stem (iPS) cell colonies from both BJ human fibroblasts and patient-derived fibroblasts1. Substitution of other media during mRNA reprogramming is not recommended, however, clonal mRNA reprogrammed iPS cell lines derived in Pluriton can be expanded and maintained in other conditions and media, such as Nutristem™ XF/FF Culture Medium. For a detailed description of the recommended usage of Pluriton for mRNA reprogramming, please see Stemgent's user manual: Stemgent mRNA Reprogramming System.
Figure 1. Comparison of Pluriton Reprogramming Medium and other common human ES culture media for iPS cell colony generation during mRNA based reprogramming. Different target cell densities (50K, 25K, or 10K per well) of BJ fibroblasts were plated on human fibroblast feeders (250K/well) in a single well of 6-well plate. Each condition wasincubated at 5% O2 and transfected with 1.2 ug of mRNA reprogramming cocktail for 16 consecutive days without enzymatic passaging. Primary cultures were assayed with the Stemgent® StainAlive™ Tra-1-81 antibody (1:100) on Day 19 and Tra-1-81 positive colonies were counted using a fluorescent microscope. Each bar in the graph is individually labeled with the number of iPS cell colonies generated.
- 500 ml Pluriton Reprogramming Medium
- 0.2 ml Pluriton Supplement 2500X
Storage and Stability
Store the Pluriton Medium at -20°C, and the Pluriton Supplement 2500X at less than -70°C. These products are stable for a minimum of 3 months when stored as directed.
Pluriton Medium and Supplement are sterile and have tested negative for mycoplasma. Pluriton mRNA Reprogramming Medium is functionally tested for successful mRNA-based reprogramming of BJ human fibroblasts. Complete reprogramming of iPS cell colonies is confirmed by continued expansion and expression of pluripotency markers.
- Stemgent mRNA Reprogramming Factors Set: hOKSML (Cat. No. 00-0067)
- Stemgent mRNA Reprogramming Kit (Cat. No. 00-0071)
- Stemgent Oct4 mRNA, Human (Cat. No. 05-0014)
- Stemgent Klf4 mRNA, Human (Cat. No. 05-0015)
- Stemgent Sox2 mRNA, Human (Cat. No. 05-0016)
- Stemgent Lin28 mRNA, Human (Cat. No. 05-0017)
- Stemgent c-Myc mRNA, Human (Cat. No. 05-0018)
- Stemgent nGFP mRNA (Cat. No. 05-0019)
- Stemgent eGFP mRNA (Cat. No. 05-0020)
- Stemfect™ RNA Transfection Kit (Cat. No. 00-0069)
Safety Data Sheets
- NuFF-RQ™ IRR (GSC3002G)
- Warren, L., Manos, P.D., Ahfeldt, T., Loh, Y.H., Li, H., Lau, F., Ebina, W., Mandal, P.K., Smith, Z.D., Meissner, A., Daley, D.Q., Brack, A.S., Collins, J.J., Cowan, C., Schlaeger, T.M., and Rossi, D.J. (2010) Highly efficient reprogramming to pluripotency and directed differentiation of human cells with synthetic modified mRNA. Cell Stem Cell 7: 618-630.
- Rajasingh S; Thangavel J; Czirok A; Samanta S. "Generation of functional cardiomyocytes from efficiently generated human iPSCs and a novel method for measuring contractility." PLoS one (2015)
- Desmarais JA; Unger C; Damjanov I; Meuth M; Andrews P. "Apoptosis and failure of checkpoint kinase 1 activation in human induced pluripotent stem cells under replication stress." Stem Cell Res Ther 7:17 (2016)
- Sridhar A; Ohlemacher SK; Langer KB; Meyer JS. "Robust differentiation of mRNA-reprogrammed human induced pluripotent stem cells toard a retinal lineage." Stem Cell Translational Medicine 5:10 (2016)
- Ohlemacher SK; Sridhar A; Xiao Y; Hochstetler AE; Sarfarazi M; Cummins TR; Meyer JS. "Stepwise differentiation of retinal ganglion cells from human pluripotent stem cells enables analysis of glaucomatous neurodegeneration." Stem Cells doi: 10.1002/stem.2356 (2016)
- Briggs SF; Dominguez AA; Chavez SL; Reijo Pera RA. "XIST in human preimplantation embryos and iPSCs." Stem Cells 33:1771 (2015)
- Durruthy JD; Sebastiano V. "Derivaton of GMP-compliant integration-free hiPSCs using modified mRNAs." Methods Mol Biol 1283: 31 (2014)
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