Rat hepatocyte spheroids formed by rocked technique maintain differentiated hepatocyte gene expression and function
| Autor: | Harris Khan, Piero Rinaldo, Scott L. Nyberg, Bruce Amiot, Rory P. Remmel, Colleen M. Brophy, Jennifer Luebke-Wheeler |
|---|---|
| Rok vydání: | 2008 |
| Předmět: |
Cell Survival
Cell Culture Techniques Gene Expression Cell Separation Biology Article law.invention Tissue culture law Cell Adhesion medicine Animals Cell adhesion Cell Shape Oligonucleotide Array Sequence Analysis Clotting factor Hepatology Spheroid Bioartificial liver device DNA In vitro Culture Media Rats Cell biology Microscopy Electron medicine.anatomical_structure Microscopy Fluorescence Cell culture Hepatocyte embryonic structures Hepatocytes RNA |
| Zdroj: | Hepatology. 49:578-586 |
| ISSN: | 0270-9139 |
| DOI: | 10.1002/hep.22674 |
| Popis: | The culture of primary hepatocytes as spheroids creates an efficient 3-dimensional tissue construct for hepatic studies in vitro. Spheroids possess structural polarity and functional bile canaliculi with normal differentiated function. Thus, hepatocyte spheroids have been proposed as the cell source in a variety of diagnostic, discovery, and therapeutic applications, such as a bioartificial liver. Using a novel rocking technique to induce spheroid formation, kinetics of spheroid formation, cell-cell adhesion, gene expression and biochemical activities of rat hepatocyte spheroids were tested over 14 days of culture. Evidence was provided that the formation of spheroids occurred faster and with fewer non-adherent hepatocytes in rocked suspension culture compared to a traditional rotational system. Hepatocyte spheroids in rocked culture showed stable expression of over 80% of 242 liver-related genes including those of albumin synthesis, urea cycle, phase I and II metabolic enzymes, and clotting factors. Biochemical activity of rocked spheroid hepatocytes was superior to monolayer culture of hepatocytes on tissue culture plastic and collagen. In conclusion, spheroid formation by rocker technique was more rapid and more efficient than rotational technique. Rocker formed spheroids appear suitable for application in a bioartificial liver or as an in vitro liver tissue construct. |
| Databáze: | OpenAIRE |
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