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Stemgent血细胞重编程试剂盒正式上市

ReproCELL旗下的血细胞重编程试剂盒,是全球第一个商业化的使用srRNA和micro RNA进行细胞重编程的试剂盒,为干细胞研究人员提供安全、灵活、快速的方法,使外周血、脐带血以及成人和新生儿的成纤维细胞中的内皮祖细胞(EPCs)重编程为iPS细胞。

 这个试剂盒主要是有三部分组成:OKSiM-srRNA,microRNA Reprogramming Cocktail 和 B18R重组蛋白。

 

 
Product Overview
 
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.
 
Product Specifications
 
Size
1 kit
 
Kit Contents
  • OKSiM-srRNA, (Part No. 05-0038), 5 µg, 1 vial
  • microRNA Reprogramming Cocktail (20 µM) (Part No. 05-0036), 35 µL, 1 vial
  • B18R Recombinant Protein, Carrier-Free (Cat. No. 03-0017), 50 µg, 1 vial
Storage and Stability
  • 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)