“#D3D Differentiation of HPSCs into Islets-like Organoids and Tissue Manufacturing”
The success in directed differentiation of human pluripotent stem cells (HPSC) including embryonic stem (hES) and induced pluripotentstem (iPS) cells into islet-like cells raises new hopes for cell-based diabetes therapy. This, however, has yet been possible due to the difficulty in generating fully functional beta cells in vitro. In many cases, cells differentiated from HPSCs are immature, or in other words, are unsuitable for cell replacement therapy. Most islet-like cells derived from HPSCs in vitro fail to function normally in vivo after transplantation in diabetic animal models. On the other hand, the in vivo maturation of pancreatic endoderm progenitors presents significant challenges. Here we report a new study on generating islets-like organoids and maturating hESCs-derived pancreatic β cells within a biomimetic 3D scaffold. We discovered that treating cells with (-)-indolactam V is critical to PDX1 and HNF6 expression divergence at Stage II of the differentiation. The expression of PDX1, not HNF6, in these cells indicated the commitment of hESCs toward pancreatic endocrine, not exocrine cell lineages. The organoids formed consisted of pancreatic α, β, d, and pancreatic polypeptide (PP) cells. A high level co-expression of PDX1, NKX6.1, and NGN3 in these cells suggests characteristics of pancreatic β cells. Most insulin-secreting cells generated did not express glucagon, somatostatin, or PP. The expression of mature β cell marker genes such as pdx1, Ngn3, Insulin, MafA, and Glut2 was detected in these cells. A high level production of C-peptide confirmed the de novo endogenous insulin production in these cells. Insulin-secretory granules were detected in these cells, further alluding their high maturity. Exposing cells to a high concentration of glucose induced a sharp increase in insulin secretion, suggesting that they are more sensitive to a glucose challenging due to their elevated maturity. The augment of this technology to other stem cell differentiations will bring cell replacement therapy one step closer to treating many diseases such as diabetes in more controllable clinical settings. In addition, emerging technologies in advanced biomanufacturing will also be discussed in the seminar.