Abstract
Background Embryonic stem (ES) cells have the potential to produce unlimited numbers of surrogate insulin-producing cells for cell
replacement therapy of type I diabetes mellitus. The impact of the in
vivo environment on mouse ES cell differentiation towards
insulin-producing cells was analysed morphologically after implantation.
Methods ES cells differentiated in vitro into insulin-producing cells
according to the Lumelsky protocol or a new four-stage differentiation
protocol were analysed morphologically before and after implantation
for gene expression by in situ reverse transcription polymerase chain
reaction and protein expression by immunohistochemistry and
ultrastructural analysis.
Results In comparison with nestin positive ES cells developed according
to the reference protocol, the number of ES cells differentiated with
the four-stage protocol increased under in vivo conditions upon
morphological analysis. The cells exhibited, in comparison to the in
vitro situation, increased gene and protein expression of Pdx1,
insulin, islet amyloid polypeptide (IAPP), the GLUT2 glucose
transporter and glucokinase, which are functional markers for
glucose-induced insulin secretion of pancreatic beta cells. Renal
sub-capsular implantation of ES cells with a higher degree of
differentiation achieved by in vitro differentiation with a four-stage
protocol enabled further significant maturation for the
beta-cell-specific markers, insulin and the co-stored IAPP as well as
the glucose recognition structures. in contrast, further in vivo
differentiation was not achieved with cells differentiated in vitro by
the reference protocol.
Conclusions A sufficient degree of in vitro differentiation is an
essential prerequisite for further substantial maturation in a
beta-cell-specific way in vivo, supported by cell-cell contacts and
vascularisation. Copyright (c) 2009 John Wiley & Sons, Ltd.
Original language | English |
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Pages (from-to) | 464-476 |
Number of pages | 13 |
Journal | Diabetes/Metabolism Research and Reviews |
Volume | 25 |
Issue number | 5 |
Early online date | 7 May 2009 |
DOIs | |
Publication status | Published - Jul 2009 |
Keywords
- amyloid
- animals
- biological markers
- cell culture techniques
- cell differentiation
- experimental diabetes mellitus
- type 1 diabetes mellitus
- embryonic stem cells
- gene expression profiling
- gene expression regulation
- glucokinase
- glucose transporter type 2
- homeodomain proteins
- insulin
- insulin-secreting cells
- intermediate filament proteins
- islet amyloid polypeptide
- male
- mice
- nerve tissue proteins
- stem cell transplantation
- trans-activators