Abstract
Pancreatic islet transplantation is the fundamental treatment for insulin-dependent diabetes; however, donor shortage is a major hurdle in its use as a standard treatment. Accordingly, differentiated insulin-producing cells (DIPCs) are being developed as a new islet source. Differentiation efficiency could be enhanced if the spheroid structure of the natural islets could be recapitulated. Here, we fabricated DIPC spheroids using concave microwells, which enabled large-scale production of spheroids of the desired size. We prepared DIPCs from human liver cells by trans-differentiation using transcription factor gene transduction. Islet-related gene expression and insulin secretion levels were higher in spheroids compared to those in single-cell DIPCs, whereas actin–myosin interactions significantly decreased. We verified actin–myosin-dependent insulin expression in single-cell DIPCs by using actin–myosin interaction inhibitors. Upon transplanting cells into the kidney capsule of diabetic mouse, blood glucose levels decreased to 200 mg/dL in spheroid-transplanted mice but not in single cell-transplanted mice. Spheroid-transplanted mice showed high engraftment efficiency in in vivo fluorescence imaging. These results demonstrated that spheroids fabricated using concave microwells enhanced the engraftment and functions of DIPCs via actin–myosin-mediated cytoskeletal changes. Our strategy potentially extends the clinical application of DIPCs for improved differentiation, glycemic control, and transplantation efficiency of islets.
Highlights
Type 1 diabetes mellitus (T1DM) is characterized by insulin deficiency that results from the destruction of pancreatic islets, a phenomenon that is the result of an autoimmune response
The sizes of differentiated insulin-producing cells (DIPCs) spheroids were measured in suspension
The spheroid size consistently increased as the cell number increased, and an association was found with DIPCs (Pearson’s coefficient: 0.922)
Summary
Type 1 diabetes mellitus (T1DM) is characterized by insulin deficiency that results from the destruction of pancreatic islets, a phenomenon that is the result of an autoimmune response (or other unknown causes). Due to the use of efficacious anti-rejection therapies and development of transplantation techniques, the long-term insulin-independence rate associated with clinical islet transplantation has improved [4,5]; donor shortage remains a major hurdle [6]. Insulin-producing cells (DIPCs)—differentiated from stem cells or adult cells—are being developed as a new islet source [7]. Somatic or adult stem cells with a non-pancreatic origin have been transdifferentiated into DIPCs directly through gene modifications [8,9,10,11]. These cells showed insulin production and genetic changes in vitro, they could not recapitulate physiological function in clinical applications
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