Book of abstracts - British Neuroscience Association

11.04
Expression profile of mesenchymal stem cells differentiated into
schwann cells
D Mahay, Shawcross S, Terenghi G
Blond McIndoe Research Laboratories,, The University of Manchester,
UK
Schwann cells (SC) are essential facilitators of peripheral nerve
regeneration following injury as they provide physical support and
guidance. In vitro these supporting cells are slow growing, hence not
well suited to a tissue engineering approach to nerve repair. We
differentiated rat bone marrow mesenchymal stem cells (MSC) into
SC-like cells using an established cocktail of growth factors. Semiquantitative
RT-PCR, Western blotting and immunocytochemistry
were used to detect glial cell marker as well as neurotrophic factors
transcripts and assess protein expression in MSC in comparison with
SC (positive control). The presence of the transcripts and proteins:
glial fibrillary acidic protein, low affinity neurotrophin receptor p75,
calcium binding protein S100 and the intermediate filament protein
nestin were investigated. Also investigated were transcripts of
neurotrophic factors: brain derived neurotrophic factor, glial derived
neurotrophic factor, ciliary neurotrophic factor, leukaemia inhibitory
factor, neurotrophin-3 and neurotrophin-4. Differentiated MSC
expressed transcripts of S100, nestin, glial derived neurotrophic factor,
leukaemia inhibitory and higher levels of brain derived neurotrophic
factor compared to SC. Likewise, MSC expressed the S100 protein
and higher protein levels of brain derived neurotrophic factor in
comparison to SC. Also, MSC and SC immunostained positively for
S100, glial fibrillary acidic protein, brain derived neurotrophic factor
and glial derived neurotrophic factor. Thus, the results indicate MSC
differentiated into SC-like cells appear to have similar morphological
and phenotypic characteristics to SC.
11.05
Adipose-derived stem cells differentiated into a schwann cell
phenotype promote neurite outgrowth in vitro
Kingham P J, Kalbermatten D F, Terenghi G
(1,2,3) Blond McIndoe Research Laboratory, The University of Manchester,
UK, (2) Department of Plastic, Reconstructive and Aesthetic Surgery,
University Hospital Basel, Switzerland
Peripheral nerve repair can be enhanced experimentally by transplanting
Schwann cells at the site of injury. The clinical application of this therapy is
however limited by donor site morbidity and the inability to generate a
sufficient number of cells in a short space of time. As an alternative, we
have investigated whether adult stem cells, isolated from adipose tissue,
can be differentiated into functional Schwann cells. Rat visceral fat was
enzymatically digested to yield rapidly proliferating fibroblast-like cells, of
which approximately 10% were positive for the stem cell markers, stro-1
and nestin. When the cells were treated with a mixture of glial growth
factors (GGF-2, bFGF, PDGF and forskolin) they adopted a bipolar
morphology similar to Schwann cells. Immunocytochemical staining
indicated that these cells also expressed the glial markers, S100 and
GFAP. In order to assess the function of these cells they were co-cultured
with the NG108-15 motor neuron-like cell line. Measurements of neurite
outgrowth were made using a computerised image analysis programme.
Differentiated stem cells released soluble factors which significantly
increased 3-fold the number of neurites expressed per NG108-15 cell
(p