[Show abstract][Hide abstract] ABSTRACT: Hepatocytes were cultured on a honeycomb-patterned polymer film (honeycomb film) formed by self-organization in order to investigate the influence of the honeycomb pattern on cell behavior. The changes in cell morphologies and actin filaments were observed by optical, fluorescence, and scanning electron microscopy. Hepatocytes were flattened, and the actin filaments appeared conspicuously in the spreading regions on a flat film. In contrast, the hepatocytes that were cultured on the honeycomb film were observed to form a spherical shape, and the actin filaments were localized inside the edge of the spheroid. The spheroids were observed within several hours after seeding on the honeycomb film; they were attached and the spheroid shape was maintained without any deformation. The spheroids expressed a higher level of liver specific function than the cell monolayers on the flat film. These results suggest that the honeycomb film is a suitable material for tissue engineering scaffolds and biomedical devices.
No preview · Article · Aug 2006 · Colloids and Surfaces A Physicochemical and Engineering Aspects
[Show abstract][Hide abstract] ABSTRACT: Fabrication and surface modification of a cell scaffold are very important in tissue engineering. In this study, the effects of pore size and surface wettability on hepatocyte adhesion and morphology was studied, using the porous poly-ε-caprolactone (PCL) film prepared by a simple casting technique. The degree of spreading of hepatocyte was enhanced with the decrease of the pore size (~10 μm). Hepatocyte motility and spreading were restricted on the patterned film having cellular-sized pores (ca. 20 μm). Partial hydrolysis of the film surface enhanced the migration and spheroid formation of the cultured hepatocyte.
No preview · Article · Oct 2002 · International Journal of Nanoscience
[Show abstract][Hide abstract] ABSTRACT: Hepatocyte spheroids are expected to be the main component of the artificial liver bioreactor for their higher function. The preparation of hepatocyte spheroids, however, can require as many as 24 to 96 h. To reduce this time, we investigated a method employing a new technique of rat hepatocyte preparation and a dynamic culture. The modified Seglen's method for standard hepatocyte isolation was altered by elimination of ethyleneglycol bis(aminoethylether) tetraacetate from the first perfusate and calcium from the second perfusate. Isolated hepatocytes were cultured in a spinner flask by spinning at 120 rpm. The modified Seglen's method was used as a control. Cells obtained by the new method were more cohesive and formed a higher proportion of cell aggregates than control cells. In the spinning culture, hepatocytes had a tendency to aggregate and 80% of cells formed spheroids within 6 h of culturing. The mean size of spheroids was 68.5 +/- 18.5 microm. Confocal laser scanning microscopy revealed that individual spheroids contained approximately 30% of nonparenchymal cells over their surface. Using the new hepatocyte preparation method followed by a spinning culture, we were able to produce hepatocyte spheroids in as few as 6 h.
No preview · Article · Jul 2002 · Artificial Organs
[Show abstract][Hide abstract] ABSTRACT: Mixed liver cell spheroids from rats consisting of hepatocytes and nonparenchymal cells lose three-dimensional structure and
function when cultured on dishes. In order to maintain the configuration and function of the spheroids, we cultured them with
collagen gel in various conditions, such as on the surface of collagen gel (group 2), between two collagen gel layers (group
3), and within collagen gel (group 4). Spheroids cultured on a standard collagen-coated dish were used as controls (group
1). Culture was continued for 10 days. Phase-contrast microscopy revealed that the spheroids of group 1 lost their spheroidal
configuration and became a monolayer within 24 h. Group 2 spheroids also spread out to a monolayer, but thus occurred at 24
to 48h. In group 3, spheroid configuration was sustained until day 10, though slightly flattened. In group 4, the spheroid
configuration was well maintained throughout the culture period. Urea synthesis of the spheroids cultured with collagen gel
was significantly higher than in group 1 between days 1 and 3. Albumin synthesis of three experimental groups was also significantly
higher than that of group 1. Although three experimental groups showed no difference in urea synthesis, albumin production
by spheroids in groups 3 and 4 was better maintained than in group 2, even toward the end of the culture period. It is concluded
that mixed liver-cell spheroid culture within collagen gels showed better maintenance of their configuration and function.
No preview · Article · Dec 2001 · Journal of Artificial Organs