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Efficient assembly of rat hepatocyte spheroids for tissue engineering applications
F J Wu1, J R Friend, C C Hsiao
1Department of Chemical Engineering and Materials Science, University of Minnesota, Minneapolis, Minnesota 55455-0132.
Biotechnology and Bioengineering
|May 20, 1996
Summary
Primary rat hepatocytes cultured as spheroids show enhanced liver function and morphology. Spinner vessel culture efficiently forms these spheroids, crucial for bioartificial liver devices and drug metabolism studies.
Area of Science:
- Hepatocyte culture
- Cellular aggregates
- Tissue engineering
Background:
- Primary rat hepatocytes cultured as monolayers show limited liver-specific function.
- Multicellular aggregates (spheroids) demonstrate enhanced function and differentiated morphology.
- Efficient spheroid formation is key for advanced cell culture applications.
Purpose of the Study:
- To describe an efficient method for forming hepatocyte spheroids in spinner vessels.
- To evaluate the viability, function, and ultrastructure of spheroids cultured in spinner vessels.
- To assess the potential of hepatocyte spheroids for bioartificial liver devices and drug metabolism studies.
Main Methods:
- Primary rat hepatocytes were cultured as multicellular aggregates (spheroids) in spinner vessels.
- Spheroid formation efficiency, cell viability, and albumin/urea production were monitored over 6 days.
- Transmission electron microscopy (TEM) was used to examine cell-cell contacts, tight junctions, and bile canaliculus-like structures.
Main Results:
- Over 80% of inoculated hepatocytes formed spheroids within 24 hours in spinner vessels, exceeding stationary culture efficiency.
- Spheroids maintained high viability and constant rates of albumin and urea production throughout the 6-day culture period.
- TEM revealed extensive cell-cell contacts, tight junctions, and microvilli-lined channels within the spheroids.
Conclusions:
- Spinner vessel culture provides an efficient method for generating functional hepatocyte spheroids.
- Hepatocyte spheroids exhibit superior liver-specific function, morphology, and ultrastructure compared to monolayer cultures.
- These spheroids represent a promising 3D cell system for bioartificial liver devices and xenobiotic drug metabolism research.
