Neural lineage cells produced by diferentiation of adipose stem cells for regenerative therapy in canine [Células de linaje neural producidas por diferenciación de células madre adiposas para terapia regenerativa en caninos]

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Alonso S.
Assafin M.
Backes M.
Barry M.A.
Bath K.-L.
Beisker W.
Benedetti-Rossi G.
Bérard D.
Blain A.
Bode H.-J.
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Universidad de Zulia
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Stem cells facilitate nerve regeneration by producing neurotrophic factors and differentiating into any type of neural lineage. Acute and chronic pathologies of the nervous system can compromise one or more of these lineages, so regenerative therapy with allogeneic stem cells will give better results if they are predifferentiated. The objective of this work was to demonstrate that the canine adipose derived mesenchymal stem cell (cAd-MSC) isolates were transdifferentiated into neural lineage in a high percentage population, using a protocol of pre induction and induction and in a determined time. Consequently, 5-10 grames (g) of five dog fat was treated at 37o C with Type I collagenase, the pellet obtained was seeded in Dulbecco’s Modified Eagle’s Medium + 10 % Fetal Bovine Serum + penicillin / streptomycin/ amphotericin B. Cells presented in passages 2-5, cASC were characterized by evaluating their ability to differentiate in threelineages, assess their pluripotency identifying the transcription factor Oct4 expresion and evaluating the morphology by scanning electron microscopy. Differentiation to neural lineage was performed in two treatment steps: pre-induction using basicfibroblast growth factor (bFGF) and induction using 2% Dimethyl sulfoxide and 200 mM of butylated hydroxyanisole (BHA).Identification of the neural lineage (nestin+) was performed by immunohistochemistry at different hours (h) after induction, counting 400 cells. The results show that the isolates were adherent to plastic and fibroblast morphology, multipotent whendifferentiated into osteocytes, chondrocytes and adipocytes, most of them were Oct4 + (55.4% ± 1.86); induced cells were able to differentiate into nestin+ cells (70%), with neuronal and neuronalmorphology four h ost induction. It was concluded that 70% of adipose derived mesenchymal stem cells present in passages 2-5 differentiated into nestin positive cells with neural and neural morphology four h post induction, and could be used in fresh orcryopreserved cell therapy. This is the first report of differentiated canine adipose stem cell culture to neural lineage in Peru.
Este trabajo se realizó con el apoyo financiero del Consejo Nacional de Ciencia y Tecnología (CONCYTEC) mediante convenio No. 409-2012-CONCYTEC-OAJ, así como por el convenio No. 541-FIDECOM-INNOVATE PERÚ-PIMEN-2015.
Palabras clave
Diferenciación neural, Célula madre, Adiposa