Transcription Factors Analysis in Neuronal-Like Cells Derived from Adult Human Bone Marrow.

Stem cells are immature cells with self-renewal capacity and ability to differentiate into multiple cell types including bone, cartilage, fat, tendon and muscle. Transplantation of stem cells or their derivatives to the adult brain have been proposed as future therapy for neurodegenerative diseases. Parkinson’s disease (PD) is a progressive neurodegenerative disease involving the deterioration of dopaminergic neurons of the substantia nigra, a part of the midbrain. Most of the currently used drugs are based on dopamine replacement which, although it reduces symptoms is limited owing to severe side effects. Alternatively, strategies of cell therapy for the treatment of PD focused on replacing damaged neurons with cells that may improve the functioning of the damaged cell population. In this study, we demonstrate that bone marrow stromal cells isolated from human iliac crest can be induced to differentiate into neural-like cells. Human bone marrow stromal cells differentiation was induced with N2 supplements, retinoic acid, butylated hydroxyanisole, isobutylmethylxanthine and dibutyryl-cyclic-AMP. A few hours after differentiation the cells changed their phenotype to neural-like cells, spindle shaped, with cell bodies and long branches. Moreover, immunostaining demonstrated the expression of specific neuronal markers such as nestin, neurofilaments and neuron specific enolase. To define the transcription factors associated with the differentiation, we used the TranSignal Protein/DNA kit (Panomics Inc). This new array-based technology allows functional analysis of dozens of eukaryotic transcription factors. Nuclear proteins were purified and incubated with mix labeled DNA binding oligonucleotides corresponding to the consensus sequences of 96 transcription factors. The DNA probes were then dissociated from the complexes and used to hybridize the TranSignal Array. We found that the differentiated human bone marrow stromal cells induced the expression of several neuron-specific transcription factors such as GAG (glycosaminoglycan), FKHR (human forkhead in rhabdomyosarcoma) and DR-2. The activation of these transcription factors in a time course manner was evident also by electrophoretic mobility shift assay. In conclusion, we show that the induced differentiation of neuronal like cells from bone marrow is associated with activation of specific neuronal transcription factors. Our study advances the possible use of autologous transplantation as a new strategy for cell therapy in neurodegenerative diseases.