Rapid Neural Differentiation of Human Adipose Tissue-derived Stem Cells Using NGF, Forskolin and bFGF
Behnaz Esfandiari 1, Masoud Soliemani 2, Saeed Kaviani 2 and Kazem Parivar 1*1Department of Biology, Science and Research Branch, Islamic Azad university, Tehran, Iran 2Department of hematology, Medical science faculty, Tarbiate Modares university, Tehran, Iran Corresponding Authors E-mail: kazem_parivar@yahoo.com
Abstract: Neural tissue has limited capacity to regenerate. There is a critical need to explore strategies in regenerative medicine that use stem cell therapy to transplant differentiated cells into nervous system disease. Several long differentiation period techniques are now using for differentiate stem cells to neuron like cells.To evaluate a novel short time induction method for differentiate of human Adipose derived stem cells(hADSCs) to neuron like cells with bFGF, Forskolin and NGF in 8 days. Adipose stromal cells were cultured. The third passage cells were used for in vitro differentiation. Neuronal differentiation was induced by treatment of the hADSCs with basic fibroblastic growth factor(bFGF), Forskolin, Nerve Growth Factor(NGF) and 1% FBS for 8 days. Flocytometery analysis showed that ,99/4% of ADSCs will express CD105 marker,97/8% of stem cells will express CD90, 3/25% of ADSCs will express CD34 and 2/77% of cells will express CD45. hADSCs were induced to differentiate into neuron-like cells as evidenced by neural morphology and the presence of neuronal markers including Microtubule-associated protein(MAP2) and b-tubulinIII by Immunocytochemistry. RT-PCR and Real-time PCR analysis indicated the upregulation of MAP2, NSE and NFM genes. hADSCs can be readily obtained from a small amount of fat tissue with minimal injury. Our findings indicate that simultaneous presence of differentiation factors such as bFGF, Forskolin, and NGF led to rapid differentiation into neuron-like cells in 8 days.
Keywords: Adipose derived stem cells; Neuron Like Cell; Neuronal Markers Back to TOC