Bile canaliculi formation by aligning rat primary hepatocytes in a microfluidic device

Abstract

In this study, we propose a microfluidic cell culture device mimicking the microscopic structure in liver tissue called hepatic cords. The cell culture area of the device was designed to align hepatocytes in two lines in a similar way to hepatic cords. Thanks to the structural design together with a cell seeding procedure, rat primary hepatocytes were successfully aligned in two lines and cultured under perfusion condition. It is shown that aligned hepatocytes gradually self-organize and form bile canaliculi along the hepatic cord-like structure. The present technique to culture hepatocytes with functional bile canaliculi could be used as an alternative to animal testing in the field of drug discovery and toxicological studies, and also be beneficial to tissue engineering applications.

18β‐glycyrrhetinic acid inhibits hepatocellular carcinoma development by reversing hepatic stellate cell‐mediated immunosuppression in mice

“…The cells were then treated with serial dilutions of GA (10,20,40, 80 mg/mL) and the control cells were treated only with solvents for 96 hr. The cell viability was assessed using a Cell Counting Kit-8 (Beyotime) according to the manufacturer's instructions.…”

New Models for the In Vitro Study of Liver Toxicity: 3D Culture Systems and the Role of Bioreactors

“…micro-/nano-fabrication techniques, micro-electronics and micro-fluidics), has allowed the creation of models where the cellular elements are integrated into controlled microenvironments, within which, in addition to the precise definition of the spatio-temporal signals individual cells are exposed to, it is also possible to perform the continuous multi-parameter monitoring of their biological responses ("lab-on-a-chip" devices). Representative examples of this approach, applied to cultured hepatocytes, and aimed at generating functional models of liver lobules, bile canaliculi and sinusoids, have been given, respectively, by Ho et al (2006), Lee et al Nakao et al, 2011. Although structurally very complex and interesting, these models of "micro-structured tissue-like environments" have major drawbacks, that limit significantly their application: they lack, in effect, the complexity of original tissue-specific microenvironments, which are typical of the situation in vivo; moreover, they do not allow cell survival for time periods higher than several hours or days.…”

Bioengineering approaches to guide stem cell-based organogenesis

“…Miniaturized and reductionist models of several organs, including the lung (Huh et al, 2010), the liver (Nakao et al, 2011), the kidney (Jang and Suh, 2010) and the gut (Kim et al, 2012), have thus far been created. For example, the 'human gut-on-a-chip' comprises two microfluidic channels, separated by a porous membrane, which is coated with ECM proteins and lined with a human intestinal epithelial cell line (Fig.…”