Archive for August, 2013

Picture Blog: mRNA Reprogramming for Human iPSCs without B18R!

Human induced pluripotent stem cells provide a great route towards personalized medicine and high accuracy drug screening. Allele Biotech has developed the most efficient method of making human iPSCs by using enhanced mRNAs, which have been adopted by leading pharmaceutical companies for clinical trials. The effects from medium-supplementing mRNAs are robust yet transient, and highly specific compared to both miRNAs (off-targets) and small molecules (unknown targets). To repress cellular immune response to introduced RNA molecules, viral protein B18R was previously used during mRNA reprogramming.

B18R is relatively expensive and inconvenient to use because it requires pre-aliquoting and -80C storage. The protocol has recently been dramatically improved at Allele through an NIDA-funded project. In our latest reprogramming run, all we needed to do was to include mRNA complex in the supplement during medium change for just a week without the need of adding any other type of molecules (such as B18R, miRNA, or chemicals) to help the mRNA mix, unlike all other known mRNA-reprogramming protocols. This advancement can make reprogramming human fibroblasts to footprint-free and xeno-free iPSCs a routine experiment for any lab to perform.

Human R-iPSCs were created without the need of B18R, dramatically reduced the cost and inconvenience. Shown is a newly formed iPSC colony.

mRNA reprogramming used to require B18R to repress cellular immune response from repeated exposure to RNA molecules.  Allele scientists lead by Dr. Jiwu Wang further developed their technology to bypass the use of B18R or any immune repressor.

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Thursday, August 15th, 2013 iPSCs and other stem cells Comments Off on Picture Blog: mRNA Reprogramming for Human iPSCs without B18R!

Picture Blog: More Efficient Reprogramming for Creating Induced Stem Cells (iPSCs)

Researchers at Allele Biotech achieved reprogramming of human fibroblasts into iPSCs within one week by including mRNA transfection supplement in daily medium change, at “bulk conversion” efficiency, and with cells seeded at a much wider density range compared to our previous publications. These significant improvements will further facilitate high throughput, large scale iPSC production using Allele’s feeder-free, xeno-free, footprint-free reprogramming, which was already a preferred method for both clinical applications and stem cell banking. The reprogramming project is currently being funded by the NIDA/NIH.

Dr. Jiwu Wang’s team at Allele Biotech (and his sons) succeeded on turning human skin cells into stem cells in 7 days. They also used a different medium than previously reported, even without B18R. Although the cells in the 15k cell well died, they are still happy of what they accomplished in all other cell densities tested.

Human skin fibroblasts were reprogrammed into stem cells using a proprietary mRNA cocktail in just a week.

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Thursday, August 1st, 2013 iPSCs and other stem cells 1 Comment