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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. 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.

journal_name

Transplant Proc

journal_title

Transplantation proceedings

authors

Joo DJ,Kim JY,Lee JI,Jeong JH,Cho Y,Ju MK,Huh KH,Kim MS,Kim YS

doi

10.1016/j.transproceed.2010.09.090

subject

Has Abstract

pub_date

2010-12-01 00:00:00

pages

4225-7

issue

10

eissn

0041-1345

issn

1873-2623

pii

S0041-1345(10)01510-1

journal_volume

42

pub_type

杂志文章

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