Improvement of the therapeutic capacity of insulin-producing cells trans-differentiated from human liver cells using engineered cell sheet

Abstract

Background Although pancreatic islet transplantation therapy is ideal for diabetes patients, several hurdles have prevented it from becoming a standard treatment, including donor shortage and low engraftment efficacy. In this study, we prepared insulin-producing cells trans-differentiated from adult human liver cells as a new islet source. Also, cell sheet formation could improve differentiation efficiency and graft survival. Methods Liver cells were expanded in vitro and trans-differentiated to IPCs using adenovirus vectors carrying human genes for PDX1, NEUROD1, and MAFA. IPCs were seeded on temperature-responsive culture dishes to form cell sheets. Differentiation efficiency was confirmed by ß cell-specific gene expression, insulin production, and immunohistochemistry. IPC suspension was injected by portal vein (PV), and IPC sheet was transplanted on the liver surface of the diabetic nude mouse. The therapeutic effect of IPC sheet was evaluated by comparing blood glucose control, weight gain, histological evaluation, and hepatotoxicity with IPC injection group. Also, cell biodistribution was assessed by in vivo/ex vivo fluorescence image tagging. Results Insulin gene expression and protein production were significantly increased on IPC sheets compared with those in IPCs cultured on conventional culture dishes. Transplanted IPC sheets displayed significantly higher engraftment efficiency and fewer transplanted cells in other organs than injected IPCs, and also lower liver toxicity, improved blood glucose levels, and weight gain. Immunohistochemical analyses of liver tissue revealed positive staining for PDX1 and insulin at 1, 2, and 4 weeks after IPC transplantation. Conclusions In conclusion, cell sheet formation enhanced the differentiation function and maturation of IPCs in vitro. Additionally, parameters for clinical application such as distribution, therapeutic efficacy, and toxicity were favorable. The cell sheet technique may be used with IPCs derived from various cell sources in clinical applications.

Enhanced Differentiation Capacity and Transplantation Efficacy of Insulin-Producing Cell Clusters from Human iPSCs Using Permeable Nanofibrous Microwell-Arrayed Membrane for Diabetes Treatment

“…The membranes with IPC clusters were transplanted to the liver surface, subcutaneous site, and peritoneal wall of 8-week-old male mice. For the liver surface transplantation, the surface of the recipient site was scratched with a dry gauze/cotton swab before transplantation to induce adhesion as described in our previous study [27]. Similar wounds were gently created on the peritoneal wall to enhance membrane attachment.…”

Overcoming the Limitations of Stem Cell-Derived Beta Cells

“…Usually, the choice of the cell source for reprogramming is based on a common background and an availability of the cells. For pancreatic beta cells, those are the gallbladder [ 17 ], liver [ 18 , 19 , 20 ], exocrine [ 21 , 22 ] and other endocrine pancreatic cell types [ 23 ]. Reprogramming is achieved by overexpression of the genes involved in pancreas development and characterizing mature beta cells such as pancreatic and duodenal homeobox 1 (PDX1), neurogenin-3 (NGN3), member of the Maf family of transcription factors MAFA, paired box protein PAX6, paired box protein PAX4, and other transcriptional factors [ 24 , 25 , 26 ].…”