title: Manufacturing of insulin-secreting spheroids with the RIN-5F cell line using a shaking culture method.
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 transferase-mediated dUTP nick-end labeling-positive 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.
  
  
  CONCLUSIONS
  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.
authors.affiliation: The Research Institute for Transplantation, Yonsei University, Seoul, Korea.
authors.forename: D J
authors.initials: DJ
authors.lastname: Joo
authors.forename: J Y
authors.initials: JY
authors.lastname: Kim
authors.forename: J I
authors.initials: JI
authors.lastname: Lee
authors.forename: J H
authors.initials: JH
authors.lastname: Jeong
authors.forename: Y
authors.initials: Y
authors.lastname: Cho
authors.forename: M K
authors.initials: MK
authors.lastname: Ju
authors.forename: K H
authors.initials: KH
authors.lastname: Huh
authors.forename: M S
authors.initials: MS
authors.lastname: Kim
authors.forename: Y S
authors.initials: YS
authors.lastname: Kim
mesh_terms: D000818:Animals; D002460:Cell Line; D003470:Culture Media; D007328:Insulin / Q000378:metabolism*; D000078790:Insulin Secretion; D008863:Microspheres
chemical_list: D003470:Culture Media; D007328:Insulin