VIRUS-FREE HUMAN IPSCS continued a b
Figure 4 – Brightfi eld microscopy shows HFFs being reprogrammed over a 21-day time period using a single transfection of Simplicon Reprogramming RNA. After 10 days of puromy- cin selection and replating on fresh PMEF cells, colonies started to emerge at days 17–21.
displayed similar reprogramming effi ciencies, ranging from 0.5% to 1%.
Conclusion This article describes a synthetic, polycis-
tronic RNA-based technology that enables effi cient reprogramming of human somatic
Figure 5 – Alkaline phosphatase staining of day 28 reprogrammed fi broblasts (HFF and BJ) gener- ated using Simplicon RNA Reprogramming technology.
cells into induced pluripotent stem cells using a single transfection event. The Simplicon RNA reprogramming technology does not rely on lentiviral, retroviral or RNA-based viruses,
but combines the effi ciency of retroviral and lentiviral reprogramming technologies with the safety of nonviral-based reprogramming methods. The technology is also a step toward a more defi ned system for iPSC generation.
In conclusion, this RNA-based reprogramming approach has broad applicability for the effi- cient generation of hiPSCs. The technique can be used to aid disease cell-modeling studies, transdifferentiation research and eventual human cell therapy and regenerative medi- cine applications.
References 1. Yamanaka S. and Takahashi, K., Induction
of pluripotent stem cells from mouse embryonic and adult fi broblast cultures by defi ned factors. Cell 2006, 126(4), 663–76.
2. Zhu, S.; Li, W. et al. Reprogramming of human primary somatic cells by OCT4 and chemical compounds. Cell Stem Cell 2010, 7(6), 651–5.
3. Yoshioka, N.; Gros, E. et al. Efficient gener- ation of human iPSCs by a synthetic self- replicative RNA. Cell Stem Cell 2013, 13(2), 246–54.
Vi Chu is R&D manager, Nick Asbrock is prod- uct manager, and Min Lu is R&D Scientist, EMD Millipore, 28820 Single Oak Dr., Temecula, Calif. 92590, U.S.A.; tel.: 951-514-4267; e-mail:
vi.chu@
emdmillipore.com;
www.emdmillipore.com
AMERICAN LABORATORY • 44 • MARCH 2015
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