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题名:Extrinsic factors promoting in vitro differentiation of insulin-secreting cells from human adipose tissue-derived mesenchymal stem cells.
作者:DAVE SD; VANIKAR AV; TRIVEDI HL;
来源:Appl Biochem Biotechnol. 2013 Jun;170(4):962-71. doi: 10.1007/s12010-013-0250-y. [ IF= 0.00 ] ]
URL :10.1007/s12010-013-0250-y
日期:20130702
摘要:Understanding of beta cell regeneration is needed to develop new treatment
modalities in diabetes mellitus. We present our experience of glucose-sensitive
insulin-secreting mesenchymal stem cells (IS-MSC) generated and differentiated
from human adipose tissue (h-AD) with application of specific differentiation
media, sans xenogenic material. h-AD from donor abdominal wall was collected in
proliferation medium composed of alpha-Minimum Essential Media, albumin,
fibroblast-growth factor and antibiotics, minced, incubated in collagenase I at
37 degrees C with shaker and centrifuged. Supernatant and pellets were separately
cultured in proliferation medium on cell + plates at 37 degrees C with 5 % CO(2)
for 10 days. Cells were harvested, checked for viability, sterility,
quantification, flow-cytometry (CD45(-)/90(+)/73(+)), and differentiated into
insulin-expressing cells using medium composed of Dulbecco's modified eagle's
medium, gene expressing upregulators and antibiotics for 3 days. They were
studied for transcriptional factors paired box genes-6(Pax-6), islet 1
transcriptional factor (Isl-1), pancreatic and duodenal homobox-1(Pdx-1).
C-peptide and insulin were measured by chemiluminescence. IS-MSC showed presence
of all three transcriptional factors and showed rise in insulin and c-peptide
level in presence of glucose stimuli. It can be concluded that the specific
extrinsic factors used in the defined differentiation media effectively and
safely promote differentiation of glucose-sensitive insulin-secreting cells from
human adipose tissue, without any genetic modulation.

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