Introduction: This study focuses on creating two types of micro biomimetic scaffolds: natural bioscaffold derived from decellularized liver and micron-scale biodegradable polylactide scaffolds infused with collagen gels, upon which human hepatocytes derived from induced pluripotent stem (iPS) cells are grown to evaluate bioactive molecules in a rapid and multiplex approach.
Methods: Sprague-Dawley rat livers were decellularized with perfusion treatment using 1% Triton X-100 and 0.1% ammonium hydroxide. Circular 8mm natural bioscaffolds with uniform thickness of 500 ¯o;m were developed from the decellularized rat livers. Collagen infused polylactide scaffolds were made by machine bioprinting to micron-scale specifications. IPS cells derived human hepatocytes were provided by Cellular Dynamics International and recellularized on these scaffolds at a density of 106 cells/scaffold. For the two dimensional (2-D) control group, 0.5 million iPS hepatocytes were seeded in the 48-well collagen-I gel coating polystyrene plates.
Results: We have shown that iPS derived hepatocytes could survive on both natural bioscaffold and collagen infused polylactide scaffolds. Throughout the 14-day culture period, significantly higher albumin biosynthesis was detected at relatively constant rates for both types of scaffolds compared to the 2D control. Multi-layered cell attachment was also observed after 3-day in vitro culture. Along with the culture time, cell-matrix penetration, cell-cell connection, and cell-matrix connection were observed. Gene expression of CYP1A2, CYP2C9, and HMGCR were stronger in the natural bioscaffold compared to the collagen infused polylactide scaffolds on day 8 cultures.
Conclusions: In this study, we describe the development of bioscaffold systems produced either from decellularized rat livers or collagen infused porous polylactide scaffolds that can support hepatocyte growth and function. The small size of these scaffolds allows for several growth conditions to be assessed at one time allowing these matrices to be used as a high-throughput assay to evaluate the genetic and functional response to liver-specific bioactive molecules.
To cite this abstract in AMA style:Wang B, Jakus A, Wu A, Wu Y, Zhang J, Shah R, Wertheim J. Development of Micro Bioscaffold as a Multiplex System To Evaluate Bioactive Molecules That Enhance Cell Function [abstract]. Am J Transplant. 2013; 13 (suppl 5). https://atcmeetingabstracts.com/abstract/development-of-micro-bioscaffold-as-a-multiplex-system-to-evaluate-bioactive-molecules-that-enhance-cell-function/. Accessed January 21, 2021.
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