Generation of insulin-secreting cells from pancreatic acinar cells of animal models of type 1 diabetes

Abstract

We recently found that pancreatic acinar cells isolated from normal adult mouse can transdifferentiate into insulin-secreting cells in vitro. Using two different animal models of type 1 diabetes, we show here that insulin-secreting cells can also be generated from pancreatic acinar cells of rodents in the diabetic state with absolute insulin deficiency. When pancreatic acinar cells of streptozotocin-treated mice were cultured in suspension in the presence of epidermal growth factor and nicotinamide under low-serum condition, expressions of insulin genes gradually increased. In addition, expressions of other pancreatic hormones, including glucagon, somatostatin, and pancreatic polypeptide, were also induced. Analysis by the Cre/loxP-based direct cell lineage tracing system revealed that these newly made cells originated from amylase-expressing pancreatic acinar cells. Insulin secretion from the newly made cells was significantly stimulated by high glucose and other secretagogues. In addition, insulin-secreting cells were generated from pancreatic acinar cells of Komeda diabetes-prone rats, another animal model of type 1 diabetes. The present study demonstrates that insulin-secreting cells can be generated by transdifferentiation from pancreatic acinar cells of rodents in the diabetic state and further suggests that pancreatic acinar cells represent a potential source of autologous transplantable insulin-secreting cells for treatment of type 1 diabetes.

Role of Cadherin-mediated Cell-Cell Adhesion in Pancreatic Exocrine-to-Endocrine Transdifferentiation

“…The purified acinar-enriched exocrine cells were precultured in RPMI 1640 medium (Sigma) with 10% fetal calf serum for 6 h. Floating cells were then collected and replated on low cellbinding dishes (Nalge Nunc) in RPMI 1640 medium supplemented with 0.5% fetal calf serum, 20 ng/ml EGF (R&D Systems) at a density of 1 ϫ 10 4 cells/cm 2 . As found in previous studies (14,17), the total cell number decreased to ϳ25% of that in the initial preparation, and cell proliferation was rarely detected under these conditions. To inhibit spheroid formation of isolated pancreatic acinar cells, the neutralizing antibody against E-cadherin (ECCD-1: Takara) was added to the culture at a concentration of 200 g/ml.…”

“…Isolation and Culture of Pancreatic Acinar Cells-Pancreatic acinar cells were isolated from streptozotocin-injected 8-weekold male C57Bl/6 mice as described (14,17). To remove almost all the ␤-cells, we used an excessive dosage of streptozotocin (200 mg/kg, intraperitoneal) for this strain, by which more than a half of the mice die within a week without insulin treatment.…”

“…Pancreatic acinar cells can convert into ductal cells, and direct evidence for acinar-to-ductal transdifferentiation has been provided both in vitro (13,14) and in vivo (15). In addition, transdifferentiation of insulin-secreting cells from pancreatic exocrine cells has been demonstrated in vitro (14,16), and cell lineage tracing revealed the origin of the newly made insulin-secreting cells to be pancreatic acinar cells (14,17).…”

Long term effects of the implantation of autologous bone marrow mononuclear cells for type 2 diabetes mellitus

“…As a result there have been many investigations into methods of preserving or recovering islet beta-cells and attempts to alter or delay the natural course of type 2 diabetes. Previous studies have shown that bone marrow stem cells can differentiate into islet cells [12][13][14] and can reduce blood glucose in animal models rendered diabetic by the use of streptozotocin [15][16][17]. Several studies of T1DM have also shown that autologous nonmyeloablative hematopoietic stem cell transplantation along with high-dose immunosuppression resulted in prolonged insulin independence in the majority of patients [18][19][20].…”

The Quest for Tissue Stem Cells in the Pancreas and Other Organs, and their Application in Beta-Cell Replacement

“…This was achieved with cells from normal mice, and from diabetic mice [54,55] (Table 2). However, this approach was associated with lower efficiency.…”