The effect of substances known as inducers of neuronal differentiation on cultured human and mouse adipose-derived mesenchymal
stem cells (ASCs) and their fate after transplantation into the injured and ischemic mouse brains were studied. ASCs were isolated
from the human and mouse adipose tissue. Inducers of neuronal differentiation included β-mercaptoethanol, glial cell line-derived
neurotrophic factor (GNDF), brain-derived neurotrophic factor (BDNF), retinoic acid (RA), 5-azacytidine, as well as their combinations.
Three days after the induction, the phenotype of the induced cells was analyzed using immunocytochemistry and real-time PCR assay for
differential expression of specific genes. The induction efficiency was evaluated by the increased transcription of neuronal differentiation
markers: nestin, β-III-tubulin (Tub-B), microtubule-associated protein 2 (MAP2), and neuron-specific enolase (ENO2). The expression of
marker genes was tested by immunocytochemical analysis. ASC cultivation in the medium with RA or BDNF in combination with 5-
azacytidine for a week increased the mRNA and protein levels of nestin, Tub-B, and ENO2. The transplantation of induced mouse ASCs
into the mouse brain increased the lifespan of the cells relative to control uninduced cells and promoted their migration from the transplantation
site to the recipient cerebral parenchyma. The transplantation of the induced cells into the mouse brain pre-exposed to endothelin-
1 promoted a more active cell migration into the surrounding ischemic brain tissue. Thus, ASC exposure to RA or BDNF in combination
with 5-azacytidine elevated the transcription of the neuronal differentiation markers and improved the viability and integration
of ASCs grafted into the mouse brain.
Keywords: Adipose-derived stem cells, induction, BDNF, 5-azacytidine, neural differentiation, transplantation, adipose-derived mesenchymal, stem cells (ASCs), ischemic mouse brains, mouse adipose tissue
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