Generation of fully functional hepatocyte-like organoids from human induced pluripotent stem cells mixed with Endothelial Cells

• Published in: Scientific reports 2019-06, Vol.9 (1), p.8920-21, Article 8920
• Main Authors: Pettinato, Giuseppe, Lehoux, Sylvain, Ramanathan, Rajesh, Salem, Mohamed M., He, Li-Xia, Muse, Oluwatoyosi, Flaumenhaft, Robert, Thompson, Melissa T., Rouse, Emily A., Cummings, Richard D., Wen, Xuejun, Fisher, Robert A.
• Format: Article
• Language: English
• Subjects: 13; 13/1; 13/100; 13/107; 13/31; 14; 14/19; 14/34; 38/77; 631/532/2064/2158; 631/532/489; 631/61/2320; 82/80; Animal models; Cell culture; Cell Differentiation; Cell Transplantation; Coculture Techniques; Endothelial cells; Endothelial Cells - cytology; Gene expression; Hepatocytes; Hepatocytes - cytology; Humanities and Social Sciences; Humans; Induced Pluripotent Stem Cells - cytology; Liver; Liver diseases; Liver Failure - therapy; Medical schools; Microvasculature; Models, Animal; Morphology; multidisciplinary; Organoids; Organoids - cytology; Pluripotency; Proteins; Science; Science (multidisciplinary); Secretion; Stem cell transplantation; Stem cells
• ISSN: 2045-2322; 2045-2322
• DOI: 10.1038/s41598-019-45514-3

Despite advances in stem cell research, cell transplantation therapy for liver failure is impeded by a shortage of human primary hepatocytes (HPH), along with current differentiation protocol limitations. Several studies have examined the concept of co-culture of human induced pluripotent cells (hiPSCs) with various types of supporting non-parenchymal cells to attain a higher differentiation yield and to improve hepatocyte-like cell functions both in vitro and in vivo . Co-culturing hiPSCs with human endothelial cells (hECs) is a relatively new technique that requires more detailed studies. Using our 3D human embryoid bodies (hEBs) formation technology, we interlaced Human Adipose Microvascular Endothelial Cells (HAMEC) with hiPSCs, leading to a higher differentiation yield and notable improvements across a wide range of hepatic functions. We conducted a comprehensive gene and protein secretion analysis of our HLCs coagulation factors profile, showing promising results in comparison with HPH. Furthermore, a stage-specific glycomic analysis revealed that the differentiated hepatocyte-like clusters (HLCs) resemble the glycan features of a mature tissue rather than cells in culture. We tested our HLCs in animal models, where the presence of HAMEC in the clusters showed a consistently better performance compared to the hiPSCs only group in regard to persistent albumin secretion post-transplantation.