Many liver diseases and liver failure are major causes of mortality that lack curative treatments. Although liver or liver cell transplantation can restore quality life, severe shortage of donor organs gravely limits the number of benefited patients. Stem cell derived hepatocytes present a renewable source of hepatic cells for use in various clinical applications. However, because of the large size of the organ any clinical application will require a large quantity of cells. To further compound the issue, for a manufacturing practice a product process must generate large treatment doses for both economic reasons and quality control needs. One can thus envision large scale processes of cell expansion and directed differentiation for clinical applications of liver cell therapy.<br/><br/>We have previously reported the conversion of pluripotent stem cells to the hepatic lineage through sequential addition of growth factors that play critical roles in embryonic liver development. These hepatocyte-like cells express important hepatic transcripts and demonstrate functional abilities through albumin and urea synthesis. However, one of the key challenges encountered is the heterogeneity of the cell populations after differentiation resulting in lower yields of desirable cells. Therefore, a robust scalable bioprocess is desired to generate enough cells for effective use in the various clinical applications. Towards this end, we have optimized a process to expand and differentiate pluripotent stem cells towards hepatocytes using microcarrier beads, to enable ease of scale up. The cells on these beads maintain the important pluripotent markers during expansion and begin to express the desired hepatic properties upon stimulation towards the hepatic fate. The quality of hepatocyte-like cells was comparable to the static cultures in terms of hepatic transcript expression and functional maturity. Furthermore, high cell concentrations on these microcarrier beads enabled probing into the dynamics of the metabolic state of the cells. Thus, this system provides a simple scalable process for the generation of large quantities of stem-cell derived hepatocytes for potential clinical applications to cater to various applications in the near future.

Scalable hepatic differentiation of human pluripotent stem cells

249th National Meeting (2015)

The official archive and repository for all ACS Meetings & Expositions abstracts and select posters or presentations.