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, YuSeun Kim

Research output: Contribution to journalArticle

9 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 1

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Insulin
Cell Line
Insulin-Secreting Cells
Carbon Monoxide
Culture Media
Pancreas
Cultured Cells
Suspensions
Cell Culture Techniques
Enzyme-Linked Immunosorbent Assay
Serum

All Science Journal Classification (ASJC) codes

  • Surgery
  • Transplantation

Cite this

Joo, D. J. ; Kim, J. Y. ; Lee, J. I. ; Jeong, J. H. ; Cho, Y. ; Ju, M. K. ; Huh, K. H. ; Kim, M. S. ; Kim, YuSeun. / Manufacturing of insulin-secreting spheroids with the RIN-5F cell line using a shaking culture method. In: Transplantation Proceedings. 2010 ; Vol. 42, No. 10. pp. 4225-4227.
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Manufacturing of insulin-secreting spheroids with the RIN-5F cell line using a shaking culture method. / Joo, D. J.; Kim, J. Y.; Lee, J. I.; Jeong, J. H.; Cho, Y.; Ju, M. K.; Huh, K. H.; Kim, M. S.; Kim, YuSeun.

In: Transplantation Proceedings, Vol. 42, No. 10, 01.12.2010, p. 4225-4227.

Research output: Contribution to journalArticle

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T1 - Manufacturing of insulin-secreting spheroids with the RIN-5F cell line using a shaking culture method

AU - Joo, D. J.

AU - Kim, J. Y.

AU - Lee, J. I.

AU - Jeong, J. H.

AU - Cho, Y.

AU - Ju, M. K.

AU - Huh, K. H.

AU - Kim, M. S.

AU - Kim, YuSeun

PY - 2010/12/1

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N2 - 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.

AB - 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.

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