The overall goal of this project is to hasten the development of cell replacement therapies for degenerative disease and injury. Human stem cell lines promise to provide all of the cell types required for transplantation. But first there is a need to control and characterize stem cell differentiation in culture. The goal of the Phase I project is to use microarray technology to identify 'expression profiles' that uniquely identify differentiated cell types. As a model system we will use the clinically relevant human Ntera-2-teratocarcinoma cell line which differentiates into neurons. The project has two specific aims:
Aim 1 : the expression pattern of 9000 genes will be determined for both the Ntera-2 precursors and the neurons. From the group of 100-200 expected differentially expressed genes, we will designate a subset of 10-20 genes as an 'expression profile panel.' Aim 2. Clinically applicable RT-PCR methods will be developed for the rapid quantification of the expression profile panel genes. In Phase 2 of the project, the technologies will be refined for clinical assays and extended to other types of stem cells. This work will provide the basis for quality control and purification processes for generating homogeneous human cell populations for cell replacement strategies.
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