Development of a Method for Embedded-Culture of Pig Pancreatic Islet-Like Cell Clusters in Agarose Containing Maltose-Carrying Polystyrene (HEVM) and Nicotinamide

Autor: Hisako Ohgawara, Akira Kobayashi, Makiko Kawamura, Sachiyo Karibe, Qian Fu, Yasue Omori, Toshihiro Akaike#
Jazyk: angličtina
Rok vydání: 1994
Předmět:
Zdroj: Cell Transplantation, Vol 3 (1994)
Druh dokumentu: article
ISSN: 0963-6897
1555-3892
09636897
DOI: 10.1177/096368979400300112
Popis: A method for embedded-culture of islet-like cell clusters (ICCs) from neonatal pig pancreas is described. The procedure is based on a sequential treatment of the pancreas with a proteolytic enzyme and ethylenediaminetetraacetate (EDTA). During the exposure to EDTA-Dispase, small pieces of pancreas were gradually digested, with gentle stirring. Under- and over-digested pancreatic cell clusters were separated from the pancreatic fragments by filtration, and formed whole islet-like cell clusters in RPMI 1640 containing 11.0 mM d-glucose and 10% fetal bovine serum (FBS). The ICCs were embedded in agarose with or without a gel containing a random copolymer hydroxethylmethacrylatevinylbenzyl maltonamide (HEVM) of hydroxethylmethacrylate- (HEMA)-vinylbenzyl maltonamide (VMA) and nicotinamide. They remained morphologically intact and a physiological response to acute stimulation with glucose was obtained when they were embedded in the agarose containing HEVM and nicotinamide. These findings suggest that ICCs in agarose containing HEVM and nicotinamide could be a useful tool for morphological, biochemical and molecular biological studies, and also as a potential source of material for transplantation.
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