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ID# CC004

A Novel Cell/Tissue Culture System to Enhance Cell Proliferation & Induce Cell Differentiation

Drs. Christopher A. Fraker, Juan Dominguez-Bendala, Camillo Ricordi & Luca Inverardi.

Problem

Current culture methods are based on the unidirectional diffusion of oxygen through the culture medium, which invariably results in the formation of steep oxygen gradients. This problem is exacerbated when it comes to 3D cultures, which are quickly becoming the new standard for tissue engineering.

Solution

The novel system is based on the use of perfluorocarbon/silicone (PFC/Si) membranes, which act as oxygen reservoirs and allow cells and tissues grown on top to breathe as in direct contact with atmospheric air. Culturing in this way improves the rate of growth and overall size, as mass transfer limitations take much longer to appear. Furthermore, the new system improves the differentiation rate of stem cells into tissues characterized by high in vivo metabolic rates. Such outcomes could not be observed by merely increasing the oxygen concentration in the incubators. As mounting evidence indicates that stem cell differentiation is directly influenced by physical variables as well as by the appropriate maintenance of 3D cell-to-cell interactions, technologies such as this will pave the way towards new culture standards.

Competitive Advantage

Provides a more physiological mode of oxygen delivery, preventing hypoxia even in thick cellular aggregates. Promotes cell viability and function Enhances differentiation in stem cell systems where oxygen has been proven to act directly as a cell specification agent.

Applications

In vitro culture of primary tissues, stem cells and cell lines. The novel system can be easily adapted to any existing cell culture platform, from flasks to small culture inserts.

Patent Status

US and PCT Applications Pending.

Licensing Opportunity

The University of Miami is seeking a company to license a novel cell/tissue culture system designed to prevent hypoxia, enhance cell proliferation and induce stem cell differentiation.

About the Inventors

Christopher Fraker, MSc., has been with the University of Miami since 1996 working on the development of perfluorinated polymers for use in the encapsulation and immunoprotection of Islets of Langerhans. His research interests also include the use of perfluorchemicals and emulsions in the preservation of donor pancreata and the culture of islets, pancreatic precursor tissues and stem cells.

Juan Domínguez-Bendala received his Ph.D. in molecular and cell biology from the University of Edinburgh/ Roslin Institute (Scotland, UK), working under the supervision of Jim McWhir, one of the creators of Dolly the sheep. He was recruited by the Diabetes Research Institute (University of Miami) in 2001 to initiate an ambitious program on stem cell research for the treatment of type I diabetes. Dr. Domínguez-Bendala is currently the Director of the Stem Cell & Translational Research laboratory at the DRI. He is one of the pioneers of human embryonic stem (huES) cell technology at the University of Miami, and has established, in collaboration with Dr. R. Pastori, groundbreaking work on protein transduction for stem cell differentiation. His work on this field was instrumental for the award of the largest grant ever given by the American Diabetes Association ($1 million; L. Inverardi, P.I.), and was featured in national and international media.

Selected References

Ricordi C, Fraker C, Szust J, Al-Abdullah I, Poggioli R, Kirlew T, Khan A, Alejandro R..Improved human islet isolation outcome from marginal donors following addition of oxygenated perfluorocarbon to the cold-storage solution.
Transplantation. 2003 May 15;75(9):1524-7.

Fraker CA, Alejandro R, Ricordi C. Use of oxygenated perfluorocarbon toward making every pancreas count. Transplantation. 2002 Dec 27;74(12):1811-2.

Domínguez-Bendala J, Priddle H, Clarke A & McWhir, J. Elevated expression of exogenous Rad51 leads to identical increases in gene targeting frequency in murine embryonic stem (ES) cells with both functional and dysfunctional p53 genes. Experimental Cell Research 10; 286(2):298-307, 2003

Domínguez-Bendala J., Klein D., Ribeiro M., Ricordi C., Inverardi L., Pastori R. and Edlund H. TAT-mediated ngn3 protein transduction stimulates pancreatic endocrine differentiation in vitro. Diabetes 54:720-726, 2005

Fraker C., Alvarez S., Papadopoulos P., Giraldo J., Gu W., Ricordi C., Inverardi L., and Dominguez-Bendala J. Enhanced oxygenation promotes beta cell differentiation in vitro. Stem Cell Express

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