Manufacturing of insulin-secreting spheroids with the RIN-5F cell line using a shaking culture method

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

Research output: Contribution to journalArticle

10 Citations (Scopus)

Abstract

Background There have been many efforts to find methods to increase insulin production by islets or modified cells. Commercially available established cell lines can be a good source of artificial islets. We manufactured sphere-shaped cell clusters composed of insulin-secreting cells from the commercially available RIN-5F cell line. Methods To generate artificial islets with insulin-secretion functions, we used the RIN-5F cell line. When cells cultured in RPMI-1640 medium containing 10% fetal bovine serum reached near confluency, they were trypsinized for suspension culture at high density, using a horizontal shaker. The cells were maintained for 5 days under 5% CO 2 with humidification. Next, the media from the RIN cell spheroid culture was collected over 5 consecutive days to test for insulin secretion. Results Spheroids of artificial islets exhibited an oval shape with an approximate size of 94.13 ± 20.41 μm on day 5 during the shaking culture. Abnormal outgrowth of spheroids was not observed. Terminal deoxynucleotidyl transferasemediated dUTP nick-end labelingpositive cells were not detected among the overall spheroids, including the core position. Insulin secretion, measured by enzyme-linked immunosorbent assay, was well maintained in the culture media over 5 days after spheroid formation. Conclusion This result suggested that a culture method with shaking can be applied to commercially available established cell lines to generate artificial islets, which might be used for a bioartificial pancreas.

Original languageEnglish
Pages (from-to)4225-4227
Number of pages3
JournalTransplantation Proceedings
Volume42
Issue number10
DOIs
Publication statusPublished - 2010 Dec

All Science Journal Classification (ASJC) codes

  • Surgery
  • Transplantation

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