Using modern proteomic and molecular tools to identify novel MSC and hESC lineage specific membrane markers: Development of new antibodies for live cell imaging and isolation
In order to move stem cell research closer to the clinic and fully realize the potential these cells have in bettering the human condition, researchers must learn as much as possible about how these cells replicate and differentiate into adult tissue. They must be able to identify cells at specific stages in complex mixtures and have the ability to isolate them for detailed analysis. The most useful set of tools for accomplishing this are antibodies which recognize specific proteins that are expressed at certain stages in the cell's path toward the fully differentiated state. There currently exists a broadly known set of markers and antibodies that have been used in the field for some time. Many of these markers are proteins that reside inside the cell which essentially requires killing the cells to allow the antibodies to recognize their target. Up to now, there have been few concerted efforts to identify new markers and develop antibodies to them. Key to isolation of specific populations of cells for basic research, diagnostics, or therapy will be to have a robust set of antibodies that recognize specific lineage markers that reside in the cell membrane and are accessible to antibodies without killing the cells one is trying to isolate.
The project proposed in this application will use the most modern cell culture, proteomic, and molecular tools to create a workflow for identification of novel lineage-specific membrane protein markers. These markers will in turn be used to create new antibody tools that will allow identification and isolation of live cells from cultures and complex tissue preparations. These new antibodies will fill a necessary (and currently somewhat empty) toolbox for fast detection and gentle isolation of cells at specific points in differentiation both for study and eventually for diagnostic and clinical use.
The work proposed here will push the boundaries of stem cell research by providing a new set of modern tools for identification and isolation of cells at specific points as they differentiate to adult tissue. These tools will be developed by a California company known to lead the field of research reagents that will ensure rapid and efficient development and broad distribution of the new tools to the research community. By partnering California tax payer money for this development and the reagent tools powerhouse of a known California-bases reagent provider, the taxpayers can be assured that repayment of their investment in the form of agreed upon royalties on sales of these tools would be forthcoming. Further, successful development of this new set of stem cell tools in California, by Californians will increase the stature of this state as a center of expertise for stem cell research.