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Immobilization of rat hepatocytes on multiporous microcarriers with larger pores and their metabolic activity
Abstract:
We have investigated the availability of multiporous microcarriers (MCs) for immobilizing isolated rat hepatocytes, but the pore size of MCs was too small (35 microns) for hepatocyte immobilization. In this study, we immobilized isolated rat hepatocytes on MCs with larger pores, and evaluated their metabolic activity. Isolated hepatocytes were immobilized on MCs precoated with collagen by the intermittent stirring method and by aspiration, and the cell-protein content per 100 mg MCs was determined for comparison of these methods. Metabolic activity was evaluated by analyzing NH3 metabolism, urea nitrogen synthesis and glucose synthesis. The aspiration method immobilized significantly more of hepatocytes on MCs than the intermittent stirring method (p < 0.05). A stationary culture of hepatocytes immobilized on MCs showed a similar NH3 metabolism to monolayer cultured hepatocytes, and hepatocytes immobilized on MCs in a floating culture showed significantly higher NH3 metabolism than those in a stationary culture (p < 0.01). However, monolayer cultured hepatocytes showed higher glucose synthesis than hepatocytes immobilized on MCs in a stationary culture (p < 0.01). In conclusion, hepatocytes immobilized on MCs proved to be useful as a bioreactor in a hybrid artificial liver.
Insights
Researchers improved hepatocyte immobilization on larger-pore microcarriers (MCs) for artificial liver applications. The aspiration method enhanced cell attachment and metabolic activity, showing promise for bioreactor development.
Area of Science:
- Biotechnology
- Hepatology
- Biomaterials
Background:
- Previous attempts at immobilizing rat hepatocytes on microcarriers (MCs) were limited by small pore sizes (35 microns).
- Optimizing microcarrier pore size is crucial for effective hepatocyte immobilization and function.
Purpose of the Study:
- To immobilize isolated rat hepatocytes on larger-pore microcarriers.
- To evaluate the metabolic activity of immobilized hepatocytes using different methods.
- To assess the potential of immobilized hepatocytes in a hybrid artificial liver system.
Main Methods:
- Isolated rat hepatocytes were immobilized on collagen-precoated, larger-pore microcarriers using intermittent stirring and aspiration methods.
- Cell-protein content was quantified to compare immobilization efficiencies.
- Metabolic activity was assessed by measuring ammonia (NH3) metabolism, urea nitrogen synthesis, and glucose synthesis.
Main Results:
- The aspiration method resulted in significantly higher hepatocyte immobilization compared to the intermittent stirring method (p < 0.05).
- Hepatocytes in a floating culture on MCs exhibited significantly higher NH3 metabolism than those in stationary culture (p < 0.01).
- Monolayer cultured hepatocytes demonstrated superior glucose synthesis compared to stationary immobilized hepatocytes (p < 0.01).
Conclusions:
- Hepatocytes immobilized on larger-pore microcarriers show potential as a functional component in hybrid artificial liver bioreactors.
- The aspiration method is more effective for hepatocyte immobilization on microcarriers.
- Floating culture conditions enhance the metabolic activity of immobilized hepatocytes, particularly ammonia metabolism.