Technology platform to develop defined culture conditions for the derivation and expansion of hESC lines
Advances in research on human embryonic stem cells (hESCs) will have major impacts on the quality of life of millions of people with health problems such as cancer, cardiovascular disease, and neurodegenerative disorders (e.g. Alzheimer’s and Parkinson’s diseases). The California Institute of Regenerative Medicine was established to develop novel cell-based therapies to treat these and other presently incurable disorders. With their ability to develop into virtually all adult cell types, hESCs represent the “raw material” for many cell-based therapies. To realize the full potential of hESCs in regenerative medicine requires, among other things, (1) establishment of well-defined culture conditions for their growth and differentiation, (2) cost-effective protocols for their expansion, and (3) derivation of new pluripotent stem cell lines under completely defined conditions. In this grant application we propose a series of experiments and the development of a novel technology platform that will provide critical information and protocols for hESC researchers.
Previous studies on factors affecting stem cell growth have focused on only one or a few elements of the cellular microenvironment, e.g., single extracellular matrix components or growth factors. Using such approaches, the screening of large numbers of factors in many combinations would be cost-prohibitive. Furthermore, currently existing hESC lines are not suitable for therapeutic use because they have been grown in poorly defined conditions containing animal-derived products, which may harbor pathogens. The proposed research will develop a high-throughput cellular microarray screening tool that incorporates combinations of recombinant proteins, synthetic materials and nanopatterned surfaces. This tool will allow the cost-effective, concomitant screening of the effect of thousands of conditions on growth and maintenance of hESCs. We combine biochemical and physical microenvironments in a controlled manner to study the responses of hESCs to physicochemical modulations of their signaling behavior and cellular fate. The results from our studies will provide fully defined and optimized culture conditions for the derivation and expansion of new hESC lines without exposure to animal-derived products.
In summary, we will develop a comprehensive approach to elucidate the responses of hESCs to a variety of factors in the microenvironment. Application of this novel and powerful technology will lead to the definition of the optimal parameters for the control of hESC growth. In addition, this technology will facilitate research on the directed differentiation of hESC into specific mature cell types, such as neurons, cardiomyocytes, pancreatic islets, and other cells, that can be applied in the treatment of a variety of debilitating human diseases.
The rise in life expectancy of the U.S. population to over 80 years will likely lead to an increase in the number of people suffering from degenerative diseases. Current medical treatments can control, but not cure, disease such as cancer, heart diseases, Alzheimer’s, and Parkinson’s. Recent advances in the study of pluripotent stem cells have provided the opportunity to develop novel strategies involving cell replacement therapies for the treatment of many currently incurable diseases and may overcome the inadequacy of the conventional drug-based treatments. Developing cell replacement therapies requires sufficiently large numbers of clinical grade human embryonic stem cells (hESCs) that can be thoroughly tested and characterized. To address this critical need we have developed a comprehensive and cost-effective technology for the high-throughput screening of conditions that regulate hESC proliferation and differentiation. The aim of our current research is to extend the capabilities of this technology platform and apply it to systematic investigation of the physicochemical conditions controlling hESCs proliferation.
Our technology will allow the screening of thousands of well-defined parameters to select the optimal culture conditions for stem cell proliferation and also differentiation. Our systematic study on these parameters, including recombinant proteins, synthetic molecules and nanopatterned surfaces, will enable the development of cost-effective large-scale production of hESCs. In addition, our study on synthetic biopolymers and peptides will eliminate the need for animal-derived products that are currently used at early stages of hESC derivation and pose potential problems for human therapies. Our technology, which we will make freely available, will additionally benefit many other lines of scientific inquiry, such as defining growth conditions of rare adult stem cell populations and modeling the cellular basis of diseases. Thus, our proposed research is fundamental to applications of hESCs in regenerative medicine and has broad benefits to researchers with a wide spectrum of scientific interests.
Our research will not only benefit the health of Californians, but also the California economy by enhancing and generating local businesses. In addition, the outcome of this project will lead to the development of a biotechnology platform that can provide great benefits to the advancement of California biotechnology. The patents, royalties and licensing fees that result from the advances in the proposed research will provide California tax revenues. Thus, the current proposed research provides not only the essential foundation for the scientific advances in regenerative medicine to improve health and quality of life, but also potential technology advancement and financial profit for the people in California.