Regenerative approaches for congenital and acquired kidney disease have the potential to repair and restore critical cell populations, renal structures, and ultimately function of damaged kidneys. The overriding objective of the studies described in this dissertation was the development, characterization, and recellularization of a three-dimensional (3D) scaffold from rhesus monkey kidneys from different age groups. This was accomplished by optimization of decellularization methods for monkey kidney transverse sections followed by characterization of extracellular matrix (ECM) proteins and biomechanical properties. It was shown that decellularized kidney scaffolds retain expression of major ECM proteins, do not contain residual cell nuclei, and show alterations in biomechanical properties as a result of the decellularization process. Age-related differences in decellularization rate and recellularization capacity using kidney explant culture were assessed. It was found that scaffolds and renal cells from younger donors provided the best outcomes in terms of recellularization potential under the culture conditions described. Recellularization was further explored by utilizing fetal renal glomerular and tubular fractions, which demonstrated that the renal tubular fraction was more effective in recellularizing tubular structures than the glomerular fraction for recellularizing glomerular structures. Proteomics analysis of decellularized kidney scaffolds revealed the presence of several non-ECM proteins. The tissue specificity of kidney and lung scaffolds were compared and showed that the decellularized scaffolds have the ability to spatially organize cells into tissue-specific structures but do not drive tissue-specific gene expression. Lastly, recellularization of kidney scaffolds using NIH-approved human embryonic stem cells (hESC) and a dynamic culture system resulted in the generation of renal constructs that contained tubules with enzymatic activity. The research addressed in this dissertation provides key approaches to using decellularized kidney scaffolds to answer meaningful questions regarding age-related differences, tissue specificity, and tissue engineering strategies for generation of renal constructs for potential regenerative strategies.
|Advisor:||Tarantal, Alice F.|
|Commitee:||Berglund, Lars, Cherry, Simon, Leach, Kent|
|School:||University of California, Davis|
|School Location:||United States -- California|
|Source:||DAI-B 74/07(E), Dissertation Abstracts International|
|Subjects:||Cellular biology, Biomedical engineering, Developmental biology|
|Keywords:||Decellularized kidneys, Hesc, Kidney, Monkey, Renal tissue, Scaffold, Stem cells|
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