Many obstacles beset islet transplantation, particularly insufficient tissue mass. Previously, we reported production of pseudo-islets. In addition, there have been reports in which coculture with pancreatic islet and bone marrow mesenchymal stem cells (BMSCs) demonstrated positive effects on pancreatic islet function. The purpose of this study was to perform morphologic and functional evaluations of pancreatic pseudo-islets cocultured with BMSCs. Pancreatic endocrine cells (PECs) were collected with a previously reported method; bone marrow was aspirated from the rat femur. Subsequently, PECs and BMSCs cocultured at high density on low-cell-binding culture dishes kept suspended by shaking. The functionality and characteristics of the mixed cell complexes were evaluated by glucose challenge, insulin enzyme-linked immunosorbent assay, reverse-transcription polymerase chain reaction, and immunohistochemistry. Through expansion for 2 weeks in continuous culture passages, ∼1 million PECs were recovered after aggregation. They presented spherical shapes and sizes similar to naïve islets, according to phase-contrast microscopy. The spheroid aggregates of pancreatic islet cells and BMSCs showed fortified functions and maintained viability. In conclusion, PECs served as a cell source for pseudo-islets, which were both morphologically and genetically similar to naïve islets. We also suggest a manufacturing method for mixed cellular complexes from 2 different origins that can improve secretion ability and cell differentiation.
|Number of pages||4|
|Publication status||Published - 2013 Jun|
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