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Functional behavior of primary rat liver cells in a three-dimensional perfused microarray bioreactor
Mark J Powers1, Dena M Janigian, Kathryn E Wack
1Division of Biological Engineering, Biotechnology Process Engineering Center, Massachusetts Institute of Technology, 77 Massachusetts Avenue, Cambridge, MA 02139, USA.
Tissue Engineering
|August 9, 2002
Summary
Pre-aggregating liver cells into spheroids before seeding in a microfabricated bioreactor significantly enhances their long-term viability and metabolic function compared to single-cell seeding. This improved 3D cell culture method maintains high albumin secretion and urea synthesis rates.
Area of Science:
- * Tissue engineering
- * Cell biology
- * Bioreactor technology
Background:
- * Microfabricated bioreactors support perfused 3D culture of liver cells.
- * Previous work established the design and operation of such bioreactors.
- * Optimizing cell seeding protocols is crucial for maintaining cell function and structure.
Purpose of the Study:
- * To assess the functional viability of liver cells in a microfabricated bioreactor.
- * To compare the influence of different seeding protocols (single-cell vs. pre-aggregated) on cell structure and function.
- * To evaluate the performance of the bioreactor in comparison with static culture conditions.
Main Methods:
- * Primary rat hepatocytes were used for cell culture.
- * Cells were seeded either as single-cell suspensions or as pre-formed spheroidal aggregates.
- * Hepatocytes were cultured in a microfabricated perfusion bioreactor and compared to static cultures.
- * Cell function was assessed by measuring albumin secretion and urea genesis rates.
- * Morphological stability was evaluated using light and electron microscopy.
Main Results:
- * Pre-aggregation of hepatocytes for 2-3 days before seeding resulted in significantly greater functional activity and morphological stability compared to single-cell seeding.
- * In single-cell cultures, albumin secretion and urea genesis rates declined significantly over 7 days, while remaining constant in pre-aggregated cultures.
- * Albumin secretion and urea genesis rates were maintained for up to 15 days post-isolation in pre-aggregated cultures.
- * Metabolic rates in the bioreactor were an order of magnitude higher than in static cultures.
- * Microscopy confirmed viable tissue structures with tight junctions, glycogen storage, and bile canaliculi.
Conclusions:
- * Pre-aggregation of primary hepatocytes into spheroids is a critical factor for enhancing cell viability and function in microfabricated perfusion bioreactors.
- * The microfabricated bioreactor system provides a superior environment for long-term 3D liver cell culture compared to static methods.
- * This optimized approach holds promise for improved in vitro models for drug testing and liver research.