Ectopic transplantation of hepatocyte sheets fabricated with temperature‐responsive culture dishes

Kanô, Kyoko; Yamato, Masayuki; Okano, Teruo

doi:10.1111/j.1872-034x.2008.00371.x

Cited by 17 publications

(17 citation statements)

References 33 publications

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“…Hepatocytes with more than 90% viability were plated on a 35-mm temperature-responsive PIPAAm culture dish (UpCell; CellSeed, Tokyo, Japan) at a density of 8 × 10 5 cells/dish. Hepatocytes were cultured as described previously (28,31) at 37°C for the first 24 h, and then the medium was replaced daily with fresh culture medium with supplemental reagents containing 0.5 μg/ml insulin (Wako Pure Chemical Industries, Osaka, Japan) and 10 ng/ml epidermal growth factor (EGF) (Invitrogen, Carlsbad, CA) until cell sheet harvesting (9).…”

Section: Methodsmentioning

confidence: 99%

Cell Shape Regulation Based on Hepatocyte Sheet Engineering Technologies

et al. 2012

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The de novo engineering of a uniform hepatocyte sheet in vitro is considered as a novel approach for liver-directed therapeutics. Hepatocytes can be cultured on a temperature-responsive culture dishes coated with poly(N-isopropylacrylamide) (PIPAAm). Following multiple days of culturing, the hepatocytes can be easily harvested as a uniform sheet by decreasing temperature from 37°C to 20°C. By modifying the sheet harvesting protocol, we have noticed that two different forms of the hepatocyte sheets, "extended" and "shrinking," were obtained. This study describes the methods for harvesting the two different forms of sheets, and their cellular structure and hepatocyte-specific functions. To obtain an "extended sheet" form, a cluster of hepatocytes covered with a support membrane was harvested by the temperature reduction. For the "shrinking sheet" form, the hepatocyte sheet was floated after reducing the culture temperature, and the floating process allowed the sheet to shrink spontaneously. Histological analysis revealed that the hepatocytes in the extended sheet form were predominantly flat, whereas the shrinking sheet contained cuboidal shaped hepatocytes. The preservation of hepatocyte-specific ultrastructures was confirmed in both types of sheets. To investigate hepatocyte-specific functionality, the harvested hepatocyte sheets were recultured on Matrigel-coated dishes. Assessment of protein production levels and chemical metabolizing activities showed the similar functionalities for each form. In contrast, the recalculation of these values per sheet versus per square centimeter of sheet surface demonstrated that the function of the shrinking sheet was significantly higher than that of the extended sheets. This study demonstrated that the hepatocyte sheets created on the PIPAAm dish could spontaneously shrink in size, but retain their hepatocyte functionality. This type of hepatocyte sheet could be utilized for the engineering of liver tissue in limited areas that are unable to give adequate transplant space.

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Section: Methodsmentioning

confidence: 99%

Cell Shape Regulation Based on Hepatocyte Sheet Engineering Technologies

et al. 2012

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“…Moreover, function, such as urea synthetic rate, decreased below the control level (Hasirci et al, 2001). In addition to collagen-based materials, many other materials and methods are proposed (Kano et al, 2008;Katsuda et al, 2010;Torok et al, 2011), which will steadily improve tissue transplantation techniques. In the future it is expected that simpler, easier, more effective and more practical transplantation techniques will be developed.…”

Section: Hepatocytes Transplantation: Where or How Are Hepatocytes Trmentioning

confidence: 99%

A Strategy Using Pluripotent Stem Cell-Derived Hepatocytes for Stem Cell-Based Therapies

Tomotsune¹,

Sato²,

Yoshie³

et al. 2011

Advances in Regenerative Medicine

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“…One advantage of using cultured cell sheets is that they eliminate the need for scaffolds, precluding the strong inflammatory responses that are typically induced when biodegradable scaffolds are broken down. Several studies have shown the potential of this technology for reconstitution of the cornea [13] and the myocardium [14], and in hepatocyte transplantation [15] and renal tube epithelial cell transfer [16].…”

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confidence: 99%

The regeneration and augmentation of bone with injectable osteogenic cell sheet in a rat critical fracture healing model

Shimizu

Akahane

Morita

et al. 2015

Injury

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Ectopic transplantation of hepatocyte sheets fabricated with temperature‐responsive culture dishes

Abstract: These results imply the usefulness of hepatocyte sheets in ectopic transplantation, as well as the need of trophic factors for hepatocyte survival.

Cited by 17 publications

References 33 publications

Cell Shape Regulation Based on Hepatocyte Sheet Engineering Technologies

Cell Shape Regulation Based on Hepatocyte Sheet Engineering Technologies

A Strategy Using Pluripotent Stem Cell-Derived Hepatocytes for Stem Cell-Based Therapies

The regeneration and augmentation of bone with injectable osteogenic cell sheet in a rat critical fracture healing model

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