Functional Characterization Of Expandable Pancreatic Progenitor-Derived β Cells Botsi In Vitro And In Vivo

Islet transplantation is one of the important therapies to treat diabetes mellitus.However,immune rejection and serious shortage of donor sources have greatly limited the application of this method.Due to infinite proliferation and pluripotency,human pluripotent stem cells(h PSCs)show great potential in cell therapy.During the last decades,although great progress has been made in the directed differentiation of h PSCs into pancreatic β cells,large-scale and low-cost production of functional β cells is still challenging.In our previous study,expandable pancreatic progenitor(e PP)cells were obtained through chemical screening,however,whether e PP cells could be further differentiated into functional pancreatic β-like(e PP-β)cells is unclear.In this study,we efficiently differentiated e PP cells into e PP-β cells using 3D directed differentiation system.These e PP-β cells highly expressed maker genes associated with β cells maturation.RNA-seq analysis evaluated the differential gene expression between e PP and e PP-β cells,and confirmed that e PP-β cells were transcriptionally similar to primary islets and h PSC-derived β(SC-β)cells.In vitro functional experiments demonstrated that e PP-β cells possessed mature mitochondria and insulin granules,and the contents of insulin and C-peptide were comparable to those in adult islets.In addition,e PP-β cells could release insulin when stimulated by high glucose.Notably,after transplantation into STZ-induced diabetes mouse models,e PP-β cells rapidly ameliorated diabetes by releasing human insulin.In summary,our research provides a new way to achieve large-scale expansion of e PP-β cells in vitro,and is expected to solve the problem of insufficient donor resources for islet transplantation,thus promoting its clinical application in the treatment of diabetes.