mRNA Reprogramming Kit

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00-0071 1 kit Ask For Price
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mrna_data_image_one
Time-course images of the generation of human iPS cells show expected morphology of iPS cells at Day 12 and 14. Day 14 fluorescent staining of TRA-1-60(10X).
mrna_data_image_two
Time-course images of the generation of human iPS cells using the mRNA Reprogramming System. Expected morphology of iPS cells at Day 6-10 (Phase 10X).
Getting Started with the mRNA Reprogramming Kit

Product Overview

The Stemgent mRNA Reprogramming Kit, in combination with the microRNA Booster Kit (00-0073), is the fastest, safest, and most efficient method for generating integration-free, virus-free, clinically relevant human iPS cells. Generate iPS cells in less than 16 days and expand new lines in culture in as little as 3 weeks as compared to lentivirus, Sendai virus, and other methods, that can take over 20 weeks to confirm that viral remnants no longer remain. This system completely eliminates the concern of virus bio-containment and safety issues.

Fully tested and validated, the Stemgent mRNA Reprogramming Kit includes five mRNA reprogramming factors, a nuclear GF P marker for monitoring transfection efficiency, the Stemgent Pluriton™ Reprogramming Medium, B18R protein, and a reproducible, optimized protocol. This kit has been validated to successfully reprogram human BJ fibroblasts as a control cell line. The Stemgent mRNA Reprogramming Kit enables researchers greater ease-of-use with the ability to simply regulate the timing of protein expression levels and have stoichiometric control of individual reprogramming factors.

Advantages

  • Non-integrating — no risk of genomic integration
  • Faster reprogramming, colonies can emerge appear in as early as 12 days
  • Stoichiometric control of individual reprogramming factors
  • Eliminates need for multi-step passaging and screening of colonies
  • Validated protocol using normal and diseased cells
  • Hands-on training and on-site support also available

Product Specifications

SIze

1 kit

Kit Contents

  • 2 vials - Stemgent Oct4 mRNA, Human (Cat. No. 05-0014), 20µg
  • 1 vial - Stemgent Klf4 mRNA, Human (Cat. No. 05-0015), 20µg
  • 1 vial - Stemgent Sox2, Human(Cat. No. 05-0016), 20µg
  • 1 vial - Stemgent Lin28 mRNA, Human(Cat. No. 05-0017), 20µg
  • 1 vial - Stemgent c-Myc mRNA, Human(Cat. No. 05-0018), 20µg
  • 1 vial - Stemgent nGFP mRNA (Cat. No. 05-0019), 20µg
  • B18R Recombinant Protein, Carrier-Free (Cat. No. 03-0017), 50µg
  • 0.2 ml of Pluriton™ Supplement 2500X (Cat. No. 01-0016)
  • 500 ml of Pluriton™ Medium (Cat. No. 01-0015)

Storage and Stability

  1. Store the box (Part 1 of 2) containing the 7 vials of mRNA, the 1 vial of B18R, and the 1 vial of Pluriton Supplement at or below -70°C. 
  2. Store the bottle (Part 2 of 2) of Pluriton Medium at -20°C.
  3. All kit components are stable for a minimum of 3 months from date of receipt, when stored as directed.

Quality Control

The Stemgent mRNA Reprogramming Kit is functionally tested for successful mRNA-based reprogramming of BJ human fibroblasts. Complete reprogramming of iPS cell colonies is confirmed by continued expression of pluripotency markers and expansion of primary iPS colonies into stable lines. All components are sterile and have tested negative for mycoplasma.

Recommended Usage

Refer to the Stemgent Protocol “Reprogramming Human Fibroblasts using the Stemgent mRNA Reprogramming Kit” for detailed instructions on material handling and productive use of the Stemgent mRNA Reprogramming Kit.

 

Documents

Limited Use License MIT

Limited Use License iPS Academia Japan

Related Products

  • 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 Lin-28 mRNA, Human(Cat. No. 05-0017)
  • Stemgent c-Myc mRNA, Human(Cat. No. 05-0018)
  • Stemgent nGFP mRNA (Cat. No. 05-0019)
  • Stemedia™ NutriStem™ XF/FF Culture Medium (Cat. No. 01-0005)
  • Stemolecule™ Y27632 (Cat. No. 04-0012)
  • Stemfactor™ Fibroblast Growth Factor-basic (Human Recombinant) (Cat. No. 03-0002)

References

  1. Pankaj K Mandal and Derrick J Rossi. (2013) Reprogramming human fibroblasts to pluripotency using modified mRNA. Nature Protocols, 8: 568 – 582.
  2. Angel, M., and Yanik, M.F. (2010) Innate immune suppression enables frequent transfection with RNA encoding reprogramming proteins. PLoS One 5: e11756.
  3. 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.
  4. Yakubov, E., Rechavi, G., Rozenblatt, S., and Givol, D. (2010) Reprogramming of human fibroblasts to pluripotent stem cells using mRNA of four transcription factors. Biochem Biophys Res Commun. 394: 189-193.

Additional Publications



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