Viability of Liver Slices Exhibiting Absorption, Metabolism, and Elimination of Substrates in Culture Medium

Shigematsu, Akiyo; Motoji, Naomi; Momose, Yuko; Iida, Aiko; Higashi, Noriko

doi:10.1006/exmp.2000.2325

“…It has been demonstrated that shear stress can affect the functions of isolated hepatocytes, [20][21][22][23] and it can be assumed that shear stress will also play a role in liver tissue culture. Perfusion culture systems will be in general associated with this drawback, and it requires the balance between shear stress and mass transfer that our own observations here and observations from other investigators 24 that the outer zone of the cultured liver slice is better preserved morphologically than the inner zone reflect. This method could not sustain the protein marker expression in all components of the cultured liver tissue after 24 h. A slight to mediocre improvement was obtained after 24 h when the tissue was cultured in the doublesided perfusion system (i.e., both sides of the tissue were perfused with fresh culture medium).…”

mentioning

confidence: 80%

Perfusion Culture Improves the Maintenance of Cultured Liver Tissue Slices

Schumacher

¹

,

Khong²,

Chang³

et al. 2007

View full text Add to dashboard Cite

Cultured precision-cut liver tissue slices are useful for studying the metabolism and toxicity of xenobiotics in liver. They may also be used to investigate the behavior of and interaction between different cell types in an intact histo-architecture. Because cultured liver tissues undergo a loss of function and morphology because of their separation from the blood supply, we investigated changes in key protein marker expressions in parenchymal and non-parenchymal cells, as well as in the extracellular matrix (ECM) at different time points. We also compared conventional culture methods such as static and dynamic cultures with perfusion culture, which allows a continuous exchange of the culture medium. In conventional culture methods, the expression of vimentin and collagen type IV decreased after 5 h in the non-parenchymal cells and the ECM, respectively, whereas the hepatocyte nuclear factor 4 alpha (HNF4a) staining in the hepatocytes remained constant. In perfusion culture, on the other hand, vimentin, collagen type IV, and HNF4a staining were clearly detectable after 5 h. The histo-architecture obtained from perfusion culture was also more compact than those obtained from conventional culture methods. After 24 h, only the perfusion cultured sample retained protein marker expression in all components of the liver tissue. Our results suggest that, to develop improved culture techniques for liver slices, changes at the early time-points should be taken into consideration. Our results also show that culture techniques that enable a continuous exchange of the culture medium seem to be superior to static or dynamic cultures in terms of maintaining the protein expression and the histo-architecture.

show abstract

“…It has been demonstrated that shear stress can affect the functions of isolated hepatocytes, [20][21][22][23] and it can be assumed that shear stress will also play a role in liver tissue culture. Perfusion culture systems will be in general associated with this drawback, and it requires the balance between shear stress and mass transfer that our own observations here and observations from other investigators 24 that the outer zone of the cultured liver slice is better preserved morphologically than the inner zone reflect. This method could not sustain the protein marker expression in all components of the cultured liver tissue after 24 h. A slight to mediocre improvement was obtained after 24 h when the tissue was cultured in the doublesided perfusion system (i.e., both sides of the tissue were perfused with fresh culture medium).…”

Section: Improved Maintenance Of Cultured Liver Tissuementioning

confidence: 80%

Perfusion Culture Improves the Maintenance of Cultured Liver Tissue Slices

Schumacher¹,

Khong²,

Shi³

et al. 2006

View full text Add to dashboard Cite

Cultured precision-cut liver tissue slices are useful for studying the metabolism and toxicity of xenobiotics in liver. They may also be used to investigate the behavior of and interaction between different cell types in an intact histo-architecture. Because cultured liver tissues undergo a loss of function and morphology because of their separation from the blood supply, we investigated changes in key protein marker expressions in parenchymal and non-parenchymal cells, as well as in the extracellular matrix (ECM) at different time points. We also compared conventional culture methods such as static and dynamic cultures with perfusion culture, which allows a continuous exchange of the culture medium. In conventional culture methods, the expression of vimentin and collagen type IV decreased after 5 h in the non-parenchymal cells and the ECM, respectively, whereas the hepatocyte nuclear factor 4 alpha (HNF4a) staining in the hepatocytes remained constant. In perfusion culture, on the other hand, vimentin, collagen type IV, and HNF4a staining were clearly detectable after 5 h. The histo-architecture obtained from perfusion culture was also more compact than those obtained from conventional culture methods. After 24 h, only the perfusion cultured sample retained protein marker expression in all components of the liver tissue. Our results suggest that, to develop improved culture techniques for liver slices, changes at the early time-points should be taken into consideration. Our results also show that culture techniques that enable a continuous exchange of the culture medium seem to be superior to static or dynamic cultures in terms of maintaining the protein expression and the histo-architecture.

show abstract