Perfusion Culture Improves the Maintenance of Cultured Liver Tissue Slices

Abstract

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.

Optimization and comprehensive characterization of a faithful tissue culture model of the benign and malignant human prostate

“…35, 36 This is particularly important for metabolic studies, and perfusion culture systems in bioreactors have been shown to enhance survival and realistically maintain cellular metabolism. 14, 36, 37 For our purposes, changing the culture medium every day appeared to enhance prostate TSC survival over changing the culture medium every other day. There was no perceived benefit to changing the medium more than once per day (data not shown).…”

“…Changing the medium daily also contributed to increased survival of TSCs. While perfusion culture might prove superior, 37 frequent refreshment of medium is a feasible alternative when using standard culture apparati.…”

A microfluidic hepatic coculture platform for cell-based drug metabolism studies

“…There is a body of evidence suggesting that fluid flow applied to various culture systems can result in better function via a variety of means including: (1) increased mass transport [27], (2) maintenance of better enzymatic activities for long term culture of primary hepatocytes [1,15] and of cell lines [28], (3) better preservation of the viability and morphology of liver tissue slices [29], and (4) better metabolism of xenobiotics by hepatocytes [16,30–32]. In our studies, we have shown that adding the element of fluid flow to the coculture system provides resolution with regard to clearance of compounds with different clearance rates, a result which is often difficult to obtain in static culture systems [33].…”

Evaluation of a microfluidic based cell culture platform with primary human hepatocytes for the prediction of hepatic clearance in human

“…There is a body of evidence demonstrating the beneficial effects of applying hydrodynamics to various cultural systems, including maintaining stronger enzymatic activities for long-term culture of primary hepatocytes [12,13] and of cell lines [14], preserving the viability and morphology of liver tissue slices [15], and the ability to perform cellular/genomic analysis of toxicity of organ tissues due to the metabolism of xenobiotics by hepatocytes [5,6,16,17]. Bader et al [18] first reported that primary rat hepatocytes in a small-scale flat-membrane bioreactor maintained drug biotrans-formation capacity of uripidil for at least 14 days.…”