Proliferation of pancreatic endocrine cells using disaggregation-expansion- reaggregation technology in isolated rat islets

J. H. Jeong, J. I. Lee, M. K. Ju, D. J. Joo, K. H. Huh, M. S. Kim, J. Y. Kim, Y. Cho, Y. S. Kim

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9 Citations (Scopus)

Abstract

Donor scarcity is a major obstacle for clinical islet transplantation. Hence, the effective use of the limited number of available islets is necessary for successful islet transplantation. We have developed a new technology that could produce pseudo-islets. Morphologic and functional evaluation was performed to test the feasibility of using these cells for transplantation. A 3-step procedure known as disaggregation-expansion-reaggregation (DER) was employed for pseudo-islet preparation. Islets isolated from 200 to 250-g male Lewis rats by collagenase digestion were separated into single cells by trypsinization. These pancreatic endocrine cells (PECs) were expanded by serial passages in culture before being aggregated at a high cell-density in a suspended state. After DER, cells were morphologically analyzed over time, and gene expression evaluated by reverse transcriptase polymerase chain reaction (RT-PCR). Through expansion by passage for 2 weeks in continuous cultures, approximately 1 million PECs were recovered after aggregation. By phase-contrast microscopy, they presented with spherical shapes and similar sizes compared with naïve islets (50-800 μm). RT-PCR results indicated expression of insulin, glucagon, and pancreatic and duodenal homeobox gene 1, which were observed in primary isolated islets as well. The insulin secretion capacity of pseudo-islets was confirmed by enzyme-linked immunosorbent assay. In conclusion, PECs treated with DER showed potential to serve as a cell source for pseudo-islet generation after in vitro cellular expansion. These cells were both morphologically and genetically similar to naïve islets. Our new technique could be a potential method to overcome the scarcity of donor islets in the near future.

Original languageEnglish
Pages (from-to)907-910
Number of pages4
JournalTransplantation Proceedings
Volume42
Issue number3
DOIs
Publication statusPublished - 2010 Dec 1

All Science Journal Classification (ASJC) codes

  • Surgery
  • Transplantation

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