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A new three-dimensional culture system for hepatocytes using reticulated polyurethane
Hepatology (Baltimore, Md.)
|April 1, 1994
Summary
Poly-N-para-vinylbenzyl-lactonamide (PVLA)-coated reticulated polyurethane (PVLA-RPU) supports long-term rat hepatocyte culture. This PVLA-RPU system maintains liver cell viability and function, showing promise for artificial liver bioreactors.
Area of Science:
- Biomaterials Science
- Hepatology
- Tissue Engineering
Background:
- Primary rat hepatocytes require specialized support for long-term culture and function.
- Traditional cell culture methods often fail to maintain hepatocyte viability and specific functions.
- Developing effective bioreactors is crucial for liver research and potential therapeutic applications.
Purpose of the Study:
- To evaluate the efficacy of Poly-N-para-vinylbenzyl-lactonamide (PVLA)-coated reticulated polyurethane (PVLA-RPU) for long-term maintenance of primary rat hepatocytes.
- To assess the morphology, viability, and liver-specific functions of hepatocytes cultured on PVLA-RPU.
- To explore the potential of the PVLA-RPU system as a bioreactor for an artificial liver.
Main Methods:
- Primary rat hepatocytes were embedded in PVLA-RPU discs at a density of 2 x 10(7) cells/cm3.
- Hepatocyte cultures were maintained in culture medium and examined using scanning and transmission electron microscopy.
- Liver-specific functions including albumin secretion, ammonium metabolism, urea synthesis, and gluconeogenesis were assessed.
- Cell viability and function were monitored over time, including in vivo studies within rat peritoneal cavities.
Main Results:
- Hepatocytes cultured on PVLA-RPU exhibited typical morphology and formed multicellular spheroids within the material's pores.
- Intercellular spaces resembling bile canaliculi were observed between hepatocytes.
- Hepatocytes maintained key liver-specific functions (albumin secretion, ammonium metabolism, urea synthesis, gluconeogenesis) and remained viable.
- These functions were sustained for over 1 month when hepatocytes were cultured in vivo within rat peritoneal cavities.
Conclusions:
- PVLA-RPU serves as an effective scaffold for the long-term culture of primary rat hepatocytes.
- The PVLA-RPU system successfully preserves hepatocyte morphology, viability, and critical liver-specific functions.
- This biomaterial-based system demonstrates significant potential for use as a bioreactor in artificial liver applications.