Coordinated Movement of Bile Canalicular Networks Reconstructed by Rat Small Hepatocytes

Sudo, Ryo; Kohara, Hiroshi; Mitaka, Toshihiro; Ikeda, Mariko; Tanishita, Kazuo

doi:10.1007/s10439-005-1690-5

Cited by 17 publications

(10 citation statements)

References 43 publications

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“…Although freshly isolated hepatocytes retain partial BC in paired cells, they are rapidly lost over time in culture (22). SHs can reconstruct functional BC that interact with NPCs when they are cultured on collagen‐coated dishes (5, 6). The maturation of SHs often occurs in the regions where they have contact with NPCs, such as stellate cells and LECs, but LECs cannot proliferate and SHs continue to proliferate without bile canalicular formation when they are cultured on collagen gel (23).…”

Section: Discussionmentioning

confidence: 99%

“…This transcellular transport is also maintained in cultured rat hepatocytes (20). SHs also display a secretory function when the cells reconstruct BC (4–6). Thus, secreted fluorescein definitely indicates BC.…”

Section: Methodsmentioning

confidence: 99%

“…Bilirubin added to the medium is secreted into the BC and accumulates without leakage. A subsequent study showed that the BC contracted and dilated continuously in a coordinated manner (6). Thus, SHs can differentiate into MHs with membrane polarity, secretory ability, and motility.…”

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confidence: 99%

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Reconstruction of 3D stacked‐up structures by rat small hepatocytes on microporous membranes

et al. 2005

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The three-dimensional (3D) culture of hepatocytes is essential for the reconstruction of functional hepatic tissues in vitro. In the present experiment, we developed a 3D-culture method in order to reconstruct hepatic cordlike structures by stacking up two-dimensional (2D) tissues composed of rat small hepatocytes (SHs), which are hepatic progenitor cells. Pairs of membranes were prepared and the cells were separately cultured on each membrane. After the SH colonies had developed, one membrane was inverted on top of the other to form an SH bilayer. Thereafter, we investigated whether the stacked cells were organized into differentiated tissues. In the 3D stacked-up structures, bile canaliculi (BC) started to form and gradually developed into anastomosing networks. Transmission electron microscopy revealed that the SHs of the upper and lower layers adhered to one another, and that BC formed between them. Bile canalicular proteins localized on the lumina of the tubular structures. Furthermore, the cells within the structures exhibited mRNA transcription of the hepatic-differentiation markers and maintained a relatively high level of albumin secretion. We conclude that highly differentiated 3D tissues, including functional BC, can be reconstructed by stacking up layers of SHs. This 3D stacked-up culture is useful for the reconstruction of tissue-engineered livers.

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Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

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Reconstruction of 3D stacked‐up structures by rat small hepatocytes on microporous membranes

et al. 2005

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“…After cells were stacked at day 2, the cells were treated with fluorescein diacetate (FD; Sigma-Aldrich) at a final concentration of 2.5 µg/ml for 15 min (12) .…”

Section: Live Cell Imagingmentioning

confidence: 99%

Morphological and Functional Changes of Rat Hepatocytes by Vertical Cell-Cell Adhesion in Three-Dimensional Stacked-Up Culture

Sudo

Mitaka

Ikeda

et al. 2008

JBSE

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Vertical cell-cell contact is an important difference between 2D-and 3D-culture models. We hypothesized that this difference in cell-cell contacts is critical in maintaining hepatocyte functions. Here we developed a simple culture technique to investigate the effect of the vertical cell-cell contacts on morphogenesis, proliferation, and differentiation of hepatocytes. Two separate monolayers of primary rat hepatocytes were first cultured on a microporous membrane and a culture dish and one was then inverted and placed on top of the other to create a 3D-configuration. Imaging techniques revealed that hepatocytes recovered cell polarity and formed bile canaliculi in response to the addition of the vertical cell-cell contacts. Quantitative analysis revealed that the cells exhibited cuboidal cell shape with vertical cell-cell contacts and maintained differentiated functions while their proliferation activity was inhibited. In contrast, hepatocytes in a monolayer gradually flattened due to the lack of vertical cell-cell contacts, resulting in cells losing differentiated functions which correlated to the deformation of cell shape. Controlling 2D-and 3D-configurations is important in switching growth and differentiation of hepatocytes. These results implicate how we apply 2D-and 3D-cultures for tissue engineering of the liver.

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“…Microvilli in bile canaliculus epithelial cells consist of Factin microfilaments, which are very important for bile excretion [7] . The continuous assembly and disassembly of Factin microfilaments promote bile duct contraction and bile excretion; therefore, the loss of Factin microfilaments leads to bile secretion disorder [9,10] . Some researchers have confirmed that cofilin and heat shock protein 27 (HSP27) are very important in the assembly and disassembly of F-actin microfilaments [11,12] .…”

Section: Introductionmentioning

confidence: 99%

Hepatic artery bridging lessens temporary ischemic injury to bile canaliculi

Wang

Liu

Wang

et al. 2015

WJG

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Author contributions: Li YM conceived the research and critically reviewed and revised the manuscript; Wang JZ and Liu Y collected and tested the specimens, and performed data analysis; all authors participated in the operations; Wang JZ drafted the manuscript; and Li YM approved the final version of the manuscript.Supported by National Natural Science Foundation of China, No. 81170454.Institutional review board statement: The study was reviewed and approved by the Second Affiliated Hospital, Xi'an Jiaotong University School of Medicine Institutional Review Board.Institutional animal care and use committee statement: All procedures involving animals were reviewed and approved by the Institutional Animal Care and Use Committee of the Xi'an Jiaotong University (IACUC protocol number: XJTULAC20120026).Conflict-of-interest statement: All authors declare that there are no conflicting interests (including but not limited to commercial, personal, political, intellectual, or religious interests) related to the work. Abstract AIM: To study whether transfer of blood between the right gastroepiploic artery and gastroduodenal artery could lessens the damage to bile canaliculi. METHODS:Forty male Bama miniature pigs were divided into four groups as follows: a control group, two hepatic artery ischemia groups (1 h and 2 h), and a hepatic artery bridging group. The hemodynamics of the hepatic artery in the hepatic artery bridging group was measured using color Doppler ultrasound. Morphological changes in the bile canaliculus were observed by transmission electron microscopy. Cofilin, heat shock protein 27 and F-actin expression was detected by immunohistochemistry, Western blot, and real-time polymerase chain reaction. Terminal deoxynucleotidyl transferase-mediated nick end-labeling method was used to evaluate liver injury. RESULTS:The hemodynamics was not changed in the hepatic artery bridging group. The microvilli in the bile canaliculus were impaired in the two hepatic artery ischemia groups. The down-regulation of cofilin and F-actin and up-regulation of heat shock protein 27 were observed in the two hepatic artery ischemia groups, while there were no significant differences between the Basic Study ORIGINAL ARTICLEcontrol group and hepatic artery bridging group. CONCLUSION:Hepatic artery ischemia aggravates damage to bile canaliculi, and this damage can be diminished by a hepatic artery bridging duct.

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Coordinated Movement of Bile Canalicular Networks Reconstructed by Rat Small Hepatocytes

Cited by 17 publications

References 43 publications

Reconstruction of 3D stacked‐up structures by rat small hepatocytes on microporous membranes

Reconstruction of 3D stacked‐up structures by rat small hepatocytes on microporous membranes

Morphological and Functional Changes of Rat Hepatocytes by Vertical Cell-Cell Adhesion in Three-Dimensional Stacked-Up Culture

Hepatic artery bridging lessens temporary ischemic injury to bile canaliculi

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