Development of culture conditions for a functional bioartificial liver

Development of culture conditions for a functional bioartificial liver
By Postdoc  Mette Hemmingsen , DTU Nanotech

In this part of the NanoBio4Trans project the task is to test and optimize the conditions for culture of stem cells and various liver cells, such as hepatocytes and endothelial cells, in the developed 3D perfusable hybrid scaffold with micro hydrogel deposits embedded with, for example, growth factors and other signaling molecules. The culture conditions should aim to provide an environment that allows for, for example, cell proliferation, stem cell differentiation and creation of a vascularized netwotk.  The testing and optimization of the cell culture conditions in the 3D perfusable hydrid scaffold will be done in parallel with the development and the design of the scaffold and the microfluidic system perfusing the scaffold. 

Fig 1 Differentiation of induced pluripotent stem cells to mature hepatocytes

Due to the fact that in vitro cultured primary liver cells have a very limited proliferation capacity, lose their activity within a few days of culture and the source is sparse, the idea is to create functional hepatocytes from Induced pluripotent stem cells inside the scaffold (Figure 1). To create sinusoidial-like organization of hepatocytes and endothelial cells (Figure 2A), hepatocytes are supposed to be cultured in a gel inside the scaffold with perfusable capillaries. Then endothelial cells can be cultured along the walls of the capillaries like in figure 2B, where mesenchymal stem cells (MSCs) are cultured along channels in gelatin. The culture of the artificial liver tissue (scaffold with cells) will take place by building it into a microfluidic system with integrated pumps and media reservoirs (Figure 2C).







Fig 2A) Co-culture of hepatocytes and endothelial cells within a perfusable hybrid scaffold with embedded micro hydrogel deposits containing necessary growth and differentiation factors. B) MSCs cultured in channels casted in a gelatin hydrogel. C) Microfluidic system with perfusable cell culture chambers.

Partners involved in the project:
25 JULY 2024