The feasibility of producing high throughput (HTP) epidermal tissue models and testing methodologies was demonstrated in Phase I studies. Epidermal tissues were produced in both 24- and 96-well plate formats. Protocols and reagents for HTP assessment of tissue viability and isolation of RNA from these tissues were also developed. Various quantitative assays for monitoring gene specific mRNA levels were analyzed. Of the methods tested, quantitative real-time polymerase chain reaction (PCR) was found to be the most versatile for HTP testing. Phase II research will further develop the results of Phase I. Phase I testing protocols will be optimized and the HTP technology will be expanded to a three-dimensional organotypic tracheal bronchial tissue model. The utility of the HTP EpiDerm tissue models and RNA isolation products will be demonstrated by quantitative real-time PCR analysis of mRNA expression of angiogenesis genes. In addition, the HTP tracheal bronchial model will be used to investigate the cytokine induced regulation of genes involved in airway inflammation and respiratory diseases.
The proposed HTP tissue models will find application in product safety assessment and gene regulation studies for a broad variety of pharmaceutical, chemical, and consumer product companies who need to screen a large number of candidate materials. The models will also be adaptable for other types of research involving modulation of gene expression in skin and airway epithelia.
