The drive to replace damaged tissues with "tissue engineered" constructs has led to fundamental questions regarding the importance of cell-cell and cell-substrate interactions in achieving the desired result. Photolithographic techniques coupled with standard silane chemistry can be readily adapted to reproducibly create arrays of cells on glass substrates, allowing control over the cell-cell and cell-substrate interactions of interest. These techniques have applications in bioartificial organs, in particular, and this study focuses on the utility of microfabrication in optimization of a bioartificial liver device.
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Editor's Preface. Foreword. Abstract. Acknowledgements. Contents. List of Figures. Introduction. 1. Tissue Engineering. 2. What Does The Liver Do? 3. What Happens When The Liver Fails? 4. Hepatic Tissue Engineering. 5. How Can Hepatocytes Be Stabilized In Vitro? 6. How Are Cell-Cell Interactions Important In Vivo? 7. Co-Culture. 8. Previous Attempts to Control Cell-Cell Interactions. 9. Micropatterning of Cells. 10. Scope of this Study. Methodology for Fabrication, Characterization, and Analysis of Micropatterned Co-Cultures. 1. Overview. 2. Fabrication of Micropatterned Co-Cultures. 3. Surface Characterization of Substrates. 4. Functional Analysis of Micropatterned Co-Cultures. 5. Mechanistic Studies. 6. Optimization Studies. 7. Summary. Characterization: Microfabricated Substrates & Co-Cultures. 1. Overview. 2. Characterization of Cell-Free Substrates. 3. Characterization of Micropatterned Cultures. 4. Discussion. 5. Summary. Functional Analysis of Micropatterned Co-Cultures. 1. Overview. 2. Characterization of Initial Cell Distribution. 3. Biochemical Analysis of Liver-Specific Function. 4. Hepatocyte Function In Situ: Immunostaining of Intracellular Albumin. 5. Haptocyte Function In Situ: Bile Duct Excretion. 6. Discussion. 7. Summary and Implications. 7. Summary and Implications. Probing Mechanisms of Hepatocyte/Fibroblast Interactions. 1. Overview. 2. Effect of Homotypic Hepatocyte Interactions on Spatial Pattern of Immunostaining. 3. Use of Conditioned Media. 4. Physical Separation of Cell Populations. 5. Agitation of Co-Cultures. 6. Discussion. 7. Summary and Future Work. Optimization of Hepatic Function in Co-Cultures. 1. Overview. 2. Reduction of Fibroblast: Hepatocyte Ratio While Preserving Heterotypic Interface in Micropatterned Co-Cultures. 3. Reduction of Fibroblast: Hepatocyte Ratio Without Control of Heterotypic Interface in Conventional, Randomly-Distributed Co-Cultures. 4. Comparison of Micropatterned and Randomly-Distributed Co-Cultures. 5. Discussion. 6. Summary. Conclusions and Outlook. 1. Summary. 2. Future Directions. References. Glossary. Index.
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