Nonviral Reprogramming Genes Accelerate Formation of Neurons from Murine Embryonic Brain Cells: Synergistic Effect of Brain Derived Neurotrophic Factor Gene Therapy

Introduction: The lesser quantity products of reprogrammed stem cells and slower differentiation of stem cells into neurons have limited the advance of cell therapy in clinical applications. Neurotrophic factors BDNF, GDNF, FGF, and IGF are critical factors for further differentiation and proliferation of neuronal cells. However, the influence of the reprogramming genes on the neurotrophic factors is unclear. Methods: Murine primary embryonic brain cells were transfected with cDNA constructs combining nonviral reprogramming genes and with/without complete length cDNA constructs of these neurotrophic factors. Reprogrammed iPSCs and progressive differentiated neural cells and controls were observed using methods of imaging and quantities. Results: Our results suggested: 1) During time-courses from the transformation of iPSCs into progressively staged neuron cells, the non-viral reprogramming genes have been significantly accelerated formations of progenitor cells, neuron cells, and neuron network, respectively. 2) The non-viral reprogramming genes directly increased gene expressions of BDNF, GDNF, FGF and IGF at RNA levels. 3) cDNA BDNF plus reprogramming genes showed a robust induction of the immature neuronal marker doublecortin at the protein level. Reprogramming genes and neurotrophic factors fuel stem cells. Conclusion: This study presents a high-efficiency approach for producing non-viral reprogrammed stem cells and auxiliary differentiated neuronal cells, which would potentially apply in the future clinical applications. One methodology is to use chemical small molecules for modifying cell culture conditions [5] for accelerating high harvest generation of neuronal cells, such as retinoic acid for neuronal differentiation [6]. The second route is to apply multiple active growth factors and to identify signaling transductions for specific cell transformations and increased rates of products. Conditions, which transformed natural stem cells into neuron cells and further differentiated into dopaminergic cells, tyrosine hydroxylase (TH)-positive cells, have been determined. These active growth factors include interleukin-1beta, glial cell line-derived neurotrophic factor (GDNF), neurturin, transforming growth factorbeta3, dibutyryl-cyclic AMP [7,8], sonic hedgehog [9-11], fibroblast growth factor-8 (FGF8) [11], and brain-derived neurotrophic factor (BDNF) [12-16], and, the signaling transductions involving Bcl-XL [17], Lmx1b/Wnt [18], developmental transcription factors ASCL1, NURR1, and LMX1A [19]. The third approach is to supply spherical neural masses as a physical method for producing the high yield of dopamine neurons [20,21]. The fourth path is to use biomaterial or combining biomaterial with biological cell growth active factors coated Citation: Liu G (2015) Nonviral Reprogramming Genes Accelerate Formation of Neurons from Murine Embryonic Brain Cells: Synergistic Effect of Brain Derived Neurotrophic Factor Gene Therapy. J Cell Sci Ther 6: 230. doi:10.4172/2157-7013.1000230