ReproCELL's Stemgent®StemRNA™-SR Reprogramming Kit is the first commercial cellular reprogramming kit combining self-replicative RNA (srRNA) and microRNA technologies to provide stem cell researchers with safe, flexible, fast and cost-effective reprogramming of both endothelial progenitor cells (EPCs) derived from human peripheral and cord blood as well as from human adult and neonatal fibroblasts.
The kit is comprised of one vial each of three components: OKSiM-srRNA, microRNA Reprogramming Cocktail and B18R Recombinant Protein.
Features include:
Robust, validated protocol for reprogramming genetically stable EPCs derived from human peripheral or cord blood
Virus- and DNA-free OKSiM reprogramming factors using self-replicative RNA
Two-transfection protocol eliminates need for conditioned medium, co-transfections, small molecules and feeder cells
iPS cell colonies generated from EPCs and fibroblasts in as few as 25 days
Protocol accomodates 5 reprogramming wells in a 6-well plate format
TABLE 1. Efficient Establishment and Cellular srRNA Reprogramming of EPCs Derived from Human Blood Samples
Primary EPC establishment efficiency for peripheral blood is cumulative of both fresh blood and cryopreserved mononuclear cell (MNC) samples. Cord blood efficiency data are from cryopreserved MNCs only. Each collagen-coated T-75 flask was seeded with a minimum of 5 x 107 MNCs. The unmodified StemRNA-SR kit protocol demonostrated high patient-to-patient efficiency using a single reprogramming attempt. Typical final yields are 10-20 iPS cell colonies per well in a 6-well plate format.
Store all three kit components at or below -70 °C.
Kit components are stable for a minimum of 3 months from date of receipt when stored as directed.
Quality Control
The StemRNA™-SR Reprogramming Kit is functionally validated for successful RNA-based reprogramming of human umbilical vein endothelial cells (HUVECs). Complete reprogramming of iPS cell colonies is confirmed by expression of pluripotency markers and appropriate colony morphology. All components of the kit are sterile and have tested negative for mycoplasma.
Limited Use Licenses
Limited Use License MIT
Limited Use License iPS Academia Japan
This product also uses technology licensed from the University of California - San Diego.
Recommended Usage
For use with associated StemRNA-SR protocols for EPC derivation and generation of iPS cells.
References
1、Yoshioka N; Gros E; Li H-R; Kumar S; Deacon DC; Maron C; Muortri AR; Chi NC; Fu X; Yu BD; Dowdy SF. "Efficient Generation of Human iPSCs by a Synthetic Self-Replicative RNA." Cell Stem Cell 13:246 (2013)
2、Geti I; Ormiston M: Touhain F; Toshner M; Movassagh M; Nichols J; Mansfield W; Southwood M; Bradley A; Rana AA; Vallier L; Morrell NW. "A practical and efficient cellular substrate for the generation of induced pluripotent stem cells." Stem Cells Trans med 1:855 (2012)
3、Yoshida Y; Takahashi K; Ishisaka T; Yamanaka S. "Hypoxia Enhances the Generation of Induced Pluripotent Stem Cells." Cell Stem Cell 5:237 (2009)
