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as well as gene editing using the CRISPR/Cas9 system. The patent-pending RNA-based reprogramming technology used by the core facility has been developed by the co-directors of the core, Drs. Ganna Bilousova and Igor Kogut, together with Dr. Dennis Roop. This novel approach provides significant advantages over traditional cell reprogramming methods, such as lentiviral-, episomal-, and Sendai virus-based methods. The approach relies on the use of non-integrating, synthetic RNA to deliver reprogramming factors. The approach used inthecoreconsistentlyreprogramshumanskinfibroblasts (including adult, disease-associated, and aged/senecent fibroblasts) with ultra-high efficiency (up to 800%) and produces high-quality iPSCs. The approach is reproducible (as reported by several independent groups) and cost-effective (significantlylessexpensivethanalternativereprogramming protocols), which makes it well-suited for the generation of biorepositories of disease-associated iPSCs.
In addition to fibroblast reprogramming, the core reprograms renal epithelial cells (RECs) isolated from urine specimens using the RNA-based approach and can assist in establishing primary fibroblast and RECs lines from human specimens. The REC reprogramming protocol was optimized with generous support from the Linda Crnic Institute for Down Syndrome and a grant from the Gates Grubstake Fund awarded to the directors of the core in 2016.
The core also provides services related to the long-term storage and maintenance of generated iPSC lines and primary cell lines. The facility is equipped with top of the line Isothermal Liquid Nitrogen Freezers, a Controlled Rate Freezing System to provide uniform freezing, and utilizes specialized Freezerworks software to ensure the integrity of the cell line database.
Since its opening, the Stem Cell Biobank core has secured reprogramming and gene-editing projects for multiple investigators from the University of Colorado and universities across the United States. Investigators affiliated with the Linda Crnic Institute for Down Syndrome at the University of Colorado Anschutz Medical Campus, the EB iPS Cell Consortium, University of Colorado Boulder, Stanford University, Columbia University, and the Mayo Clinic Arizona are among the clients of the core.
Following is a brief description of our three original core facilities established in 2011 and graphic summaries illustrating their use by both Gates Center members and nonmembers.
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