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Hepatocyte Dynamics in a Three-Dimensional Rotating Bioreactor

Hepatocyte Dynamics in a Three-Dimensional Rotating Bioreactor

Mitsuo Miyazawa, Takahiro Torii, Yasuko Toshimitsu, Katsuya Okada, Isamu Koyama

J Gastroenterol Hepatol. 2007 Nov;22(11):1959-64.

PMID: 17914977

Abstract:

Background and aims:
The use of an artificial liver system with extracorporeal circulation or a three-dimensional bioreactor perfused with liquid culture medium inevitably exposes hepatocytes to fluid mechanical stress (MS). The expression of liver-specific hepatocyte functions seems to be modulated by the magnitude of MS. Nonetheless, few studies have focused on the direct effects of MS on hepatocytes. We subjected hepatocytes to MS using an MS loading device and investigated the effects on the cytoskeleton and hepatocyte dynamics inside three-dimensional scaffolds by monitoring the changes in actin fiber, one of the components of the cytoskeleton. We also assessed the influence of MS on specific hepatocyte functions.
Methods:
We subjected hepatocytes to MS by a rotating radial flow bioreactor (RRFB) and examined the effects by comparing the MS-loaded culture cells with cells cultured under stationary conditions without MS loading. The hepatocytes (1 x 10(6)/cm(3)) were seeded on gauze without collagen coating and examined to determine morphological changes after 60 h incubation. Actin filaments in samples from the MS-loaded hepatocyte culture were stained by fluorescein isothiocyanate-labeled phalloidin.
Results:
Hepatocyte aggregation was observed in the MS-loaded culture, but not in the unloaded stationary culture. Better albumin products were observed in the MS-loaded group than in the stationary culture group at all measurement points. Actin filaments extended toward the scaffold after the start of MS loading incubation and polymerized around the hepatocytes. The hepatocyte aggregation eventually advanced to the formation of spheroids.
Conclusion:
These results suggest that MS-induced polymerization of actin filaments stimulate hepatocyte aggregation and thereby improve hepatocyte-specific function.

Chemicals Related in the Paper:

Catalog Number	Product Name	Structure	CAS Number	Price
IAR424157	Phalloidin, Fluorescein Isothiocyanate Labeled			Price

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