ReproCELL Stemgent RNA Reprogramming Systems: Fibroblast Cells

00-0076 – Stemgent® StemRNA™-NM Reprogramming Kit


ReproCELL Stemgent’s latest evolution in RNA reprogramming combines non-modified RNA and microRNA technology to provide a kit for stem cell researchers with a new level of versatility, simplicity and time savings, enabling cellular reprogramming of human fibroblasts, and cells from blood and now urine for difficult to reprogram patient samples.


Key Benefits

• Flexible technology generates high-quality human iPS cell lines from multiple target cell types
Simplified protocols minimize processing steps, increasing researcher’s out-of-the-box success rate for reprogramming of human neonatal and adult fibroblasts, endothelial progenitor cells (EPCs) derived from fresh or frozen human peripheral and cord blood, and urine-derived epithelial cells (UDCs).

• Simplified protocols and high efficiency for out-of-the-box reprogramming success
Stemgent’s StemRNA-NM requires as few as four additional reagents beyond the three kit components depending upon the cell type, unlike other technologies requiring 11 or more additional reagents. Double-stranded microRNAs enhance reprogramming efficiency for the generation of stable iPS cell lines from refractory patient samples. As few as 25,000 starting cells are needed to generate industry-leading efficiencies for fibroblasts (2 – 4%), EPCs (0.4 – 3%), and UDCs (0.1 – 0.5%).

• Time-saving protocol delivers faster results facilitating higher throughput
Colonies are ready to pick in only 10-14 days for fibroblasts and 12-16 days for EPCs and UDCs. This kit eliminates the need for screening primary iPS cell colonies, saving researchers an additional 2 to 10 weeks (3 – 10+ passages) needed for vector clearance required by other reprogramming technologies.

StemRNA-NM (Cat. No 00-0076) StemRNA-SR (Cat. No. 00-0075)
Features Fibroblasts Urine (UDCs) Blood (EPCs) Blood (EPCs)
No. wells per kit 9 3 3 5
No. Transfections required 4 8 8 2
Days to primary iPS cell colonies 10-14 14-16 14-16 28
Reprogramming efficiency 2-4% 0.1-0.5% 0.4-3% 0.001-0.005%
Screening required No No No Yes
Xeno-compatible protocol Yes No No No
GMP-compatible RNA manufacturing protocol Yes Yes Yes No

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00-0076 – Stemgent® StemRNA™-NM Reprogramming Kit

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microRNA-boosted mRNA reprogramming system

The ReproCELL Stemgent microRNA-boosted mRNA reprogramming system, combining the mRNA Reprogramming Kit (00-0071) with the microRNA Booster Kit (00-0073) and the Stemfect™ RNA Transfection Kit (00-0069), provides high-quality reprogramming of adult and neonatal fibroblasts. The microRNA Booster Kit (00-0073) increases the success with highly refractory target cells, improves efficiency, and reduces the cost per well.



00-0071 – mRNA Reprogramming Kit

The Stemgent mRNA Reprogramming Kit (00-0071), 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.

00-0071 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)

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00-0073 – microRNA Booster Kit

The Stemgent microRNA Booster Kit enhances and simplifies mRNA reprogramming for fast, efficient iPS cell line generation from both normal and patient specific cell lines. The microRNA Booster Kit includes the microRNA Reprogramming Cocktail and B18R Recombinant Protein.

The microRNA-enhanced mRNA Reprogramming method improves upon the first generation mRNA Reprogramming Kit (00-0071), delivering faster reprogramming kinetics, a streamlined protocol and improved efficacy on hard-to-reprogram or refractory cell types. The microRNA-enhanced method combines Stemgent’s mRNA Reprogramming Kit with the microRNA Booster Kit and the Stemfect™ RNA Transfection Kit to generate expandable primary iPS cell colonies in as little as 8 days. This new method improves kinetics and reduces mRNA dose per transfection resulting in a per well cost reduction of approximately 35% to 45% versus feeder-based mRNA Reprogramming.

00-0073 Contents:

• 2 vials – microRNA Reprogramming Cocktail (20 µM) (Cat. No. 05-0036), 35 µL
• B18R Recombinant Protein, Carrier-Free (Cat. No. 03-0017), 50 µg

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00-0069 – Stemgent Stemfect RNA™ Transfection Kit

The Stemfect RNA Transfection Kit is a proprietary mixture of lipid components is specifically designed for in vitro RNA transfection. This kit has been shown to deliver mRNA, siRNA and miRNA. Stemfect RNA Transfection Kit provides greater than or equal to 90% transfection efficiency of mRNA into a range of cell types with greater than 95% viability; these cell types include human embryonic stem (hES) cells, Jurkat cells, human fibroblasts, and HEK293T cells. Stemfect RNA Transfection Kit has been successfully used to deliver RNA to human ES cells, dendritic cells, and retinal pigment epithelial cells. There is no need to serum-starve cells using this kit as the transfection efficiency remains greater than or equal to 90% in the presence of serum.

00-0069 Contents:

• 1 vial – Stemfect™ RNA Transfection Reagent: 750 μL
• 1 vial – Stemfect™ Transfection Buffer: 30mL

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Expert Scientific Consultation From ReproCELL Stemgent

The ability to reprogram human tissue samples into induced pluripotent stem (iPS) cells has potentially profound implications for pharmaceutical companies in the quest to develop safe and effective new drugs in a cost effective manner. However, developing the in-house capability to reproducibly and efficiently reprogram cells can take months to years to establish and can divert significant resources from your drug discovery efforts.

Our scientists can work with your lab team to provide customized training to enable them to successfully create and culture iPS cells in your lab to ensure that your success continues beyond the end of the project.

ReproCELL Stemgent’s reprogramming services team has mastered the scientific and technical challenges of reprogramming and routinely generates custom iPS cell lines. By taking advantage of Stemgent’s unique combination of state-of-the-art technology and reprogramming expertise you can overcome the technical hurdles encountered with iPS cell generation and quickly harness the full potential of iPS cell technology in drug discovery.

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