In vitro evaluation of different protocols for the induction
of mesenchymal stem cells to insulin-producing cells
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Abstract
Stem cells therapy is a new promising approach for
diabetes mellitus (DM) treatment, but the insulin secretion
rate in differentiated cells is low when compared with pancreas
beta cells embedded in Langerhans islets. In this study,
we evaluated different protocols of insulin secretion to
achieve the most appropriate protocol for in vitro insulin
secretion. We differentiated human umbilical cord matrixderived
mesenchymal cells (hUCMs) into insulinproducing
cell (IPC) by the aim of three previously reported
protocols and a modified protocol. The insulin content was
analyzed through gene expression and immunocytochemistry
(IHC). Dithizone (DTZ) staining was done for identification
of islet-like structures. Insulin and C peptide secretion
was measured by chemiluminesence immunoassay (CLIA)
and enzyme immunoassay (EIA) as well. Reverse
transcription-PCR (RT-PCR) showed efficient expression of
insulin genes in all the study groups. IHC analysis showed
higher expression of insulin and proinsulin proteins in the
modified protocol. DTZ staining exhibited variable islet-like
clusters in the different protocols except control. This finding
was confirmed by the higher response to glucose challenge
test in this group. A modified protocol using an intermediate
step that makes the cells vulnerable to nestin production
in combination with inducing agent results in the
higher differentiation of stem cells into insulin-producing
cells and more insulin secretion in vitro