Comparison of the transcriptomic profile of hepatic human induced pluripotent stem like cells cultured in plates and in a 3D microscale dynamic environment

Abstract

We have compared the transcriptomic profiles of human induced pluripotent stem cells after their differentiation in hepatocytes like cells in plates and microfluidic biochips. The biochips provided a 3D and dynamic support during the cell differentiation when compared to the 2D static cultures in plates. The microarray have demonstrated the up regulation of important pathway related to liver development and maturation during the culture in biochips. Furthermore, the results of the transcriptomic profile, coupled with immunostaining, and RTqPCR analysis have shown typical biomarkers illustrating the presence of responders of biliary like cells, hepatocytes like cells, and endothelial like cells. However, the overall tissue still presented characteristic of immature and foetal patterns. Nevertheless, the biochip culture provided a specific micro-environment in which a complex multicellular differentiation toward liver could be oriented.

Optimized protocol for the hepatic differentiation of induced pluripotent stem cells in a fluidic microenvironment

“…In our previous studies, we showed that by applying common protocols of differentiation to microfluidic biochips, function was enhanced. However, those results were still found to be closer to those obtained with fetal cells than those that could be expected from the culture of mature primary hepatocytes (Leclerc et al, 2017). In this study, we showed that one of the critical points for the efficient culture of hiPSCs-derived hepatocytes-like cells was the preservation of the function of hepatic progenitors during the transfer in the biochips.…”

“…While the values were in a higher range than our previous report in microfluidic biochips (Leclerc et al, 2017), the tissue still exhibited primitiveness patterns. A production of albumin comparable to previous HepG2 reports in microfluidic biochips was observed.…”

“…While further improvement of the protocol could be performed in the form of cell selection (Kido et al, 2015), we here investigated the improvement of the maturation into hepatocytes by transferring the cells in a microfluidic culture system. However, those results were still found to be closer to those obtained with fetal cells than those that could be expected from the culture of mature primary hepatocytes (Leclerc et al, 2017). However, those results were still found to be closer to those obtained with fetal cells than those that could be expected from the culture of mature primary hepatocytes (Leclerc et al, 2017).…”

“…The design and fabrication process reported in our previous work (Leclerc et al, 2017) was used. The biochips were fabricated by replica moulding in polydimethylsiloxane.…”

Pluripotent stem cells to hepatocytes, the journey so far

“…A key to the success of these protocols is the ability to generate definitive endoderm cells with high efficiency, mainly by using activin A and Wnt3A (47,48), and this approach has yielded functionally improved hepatocyte-like cells. Efforts to differentiate iPS cells under 3D dynamic environments also helped to enhance the function, but were also more like foetal hepatocytes and lacked maturity (49).…”