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题名:Effect of substrate rigidity in tissue culture on the function of insulin-secreting INS-1E cells.
作者:NAUJOK O; BANDOU Y; SHIKAMA Y; FUNAKI M; LENZEN S;
来源:J Tissue Eng Regen Med. 2014 Jan 8. doi: 10.1002/term.1857. [ IF= 0.00 ] ]
URL :10.1002/term.1857
日期:20140217
摘要:Insulin-secreting INS-1E cells are a useful tool in diabetes research. However,
during permanent culture the cells tend to lose their beta cell phenotype, with
resultant loss of insulin-secretory responsiveness. This can be at least
partially attributed to inappropriate cell culture conditions. One of the
important causative factors is the rigidity of the extracellular matrix. We have
therefore systematically studied the performance of INS-1E insulin-secreting
cells cultured on polyacrylamide gels of different stiffnesses and analysed
changes in insulin content and secretion, glucokinase enzyme activity, gene
expression of beta cell transcription factors and cell death and proliferation
rates. INS-1E cells were cultured on polyacrylamide gels with a wide range of
rigidities, including the one that simulates the stiffness of the pancreas. We
detected changes in insulin content and the insulin-secretory response to glucose
stimulation in parallel to the increasing stiffness of the polyacrylamide gels in
the range 1700-111 000 Pa. On substrates with the highest and lowest rigidities,
322 and 111 000 Pa, the cells mainly formed pseudo-islets, while at rigidities of
1700-64800 Pa, including the rigidity of native pancreas tissue (3100 Pa), cells
grew as a monolayer attached to the polyacrylamide gel surface. These
observations provide evidence for an apparent mechanosensitivity of
insulin-secreting INS-1E cells affecting morphology and cellular functions. The
results can also provide practical advice regarding a selection of the materials
appropriate for successful cell culture of insulin-secreting cells. Copyright (c)
2014 John Wiley & Sons, Ltd.

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