title: Effects of nerve growth factor and basic fibroblast growth factor dual gene modification on rat bone marrow mesenchymal stem cell differentiation into neuron-like cells in vitro.
abstract: Recent studies regarding regenerative medicine have focused on bone marrow mesenchymal stem cells (BMSCs), which have the potential to undergo neural differentiation, and may be transfected with specific genes. BMSCs can differentiate into neuron‑like cells in certain neurotropic circumstances in vitro. Basic fibroblast growth factor (bFGF) and nerve growth factor (NGF) are often used to induce neural differentiation in BMSCs in vitro. However, previous studies regarding their combined actions are insufficient. The present study is the first, to the best of our knowledge, to thoroughly assess the enhancement of neural differentiation of BMSCs following transfection with bFGF and NGF. Sprague‑Dawley (SD) rat BMSCs were separated through whole bone marrow adherence, and were then passaged to the third generation. The cells were subsequently divided into five groups: The control group, which consisted of untransfected BMSCs; the plv‑blank‑transfected BMSCs group; the plv‑bFGF‑transfected BMSCs group; the plv‑NGF‑transfected BMSCs group; and the plv‑NGF‑bFGF co‑transfected BMSCs group. Cell neural differentiation was characterized in terms of stem cell molecular expression, and the neuronal morphology and expression of neural‑like molecules was detected in each of the groups. A total of 72 h post‑transfection, the expression levels of neuron‑specific enolase, glial fibrillary acidic protein, and nestin protein, were higher in the co‑transfected group, as compared with the other groups, the expression levels of β‑tubulin III were also increased in the co‑transfected cells, thus suggesting the maturation of differentiated neuron‑like cells. Furthermore, higher neuronal proliferation was observed in the co‑transfected group, as compared with the other groups at passages 2, 4, 6 and 8. Western blotting demonstrated that the transfected groups exhibited a simultaneous increase in phosphorylation of the AKT and extracellular signal‑regulated kinases (ERK) signaling pathway. These results suggested that manipulation of the ERK and AKT signaling pathway may be associated with the differentiation of transfected BMSCs.
authors.affiliation: Department of Orthopedics, The First Hospital Affiliated to Dalian Medical University, Dalian, Liaoning 116011, P.R. China.
authors.forename: Yang
authors.initials: Y
authors.lastname: Hu
authors.affiliation: Institute of Cancer Stem Cells, Cancer Center, Dalian Medical University, Dalian, Liaoning 116044, P.R. China.
authors.forename: Yan
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: Department of Orthopedics, The First Hospital Affiliated to Dalian Medical University, Dalian, Liaoning 116011, P.R. China.
authors.forename: Kang
authors.initials: K
authors.lastname: Tian
authors.affiliation: Department of Orthopedics, The First Hospital Affiliated to Dalian Medical University, Dalian, Liaoning 116011, P.R. China.
authors.forename: Chong
authors.initials: C
authors.lastname: Xun
authors.affiliation: Department of Orthopedics, The First Hospital Affiliated to Dalian Medical University, Dalian, Liaoning 116011, P.R. China.
authors.forename: Shouyu
authors.initials: S
authors.lastname: Wang
authors.affiliation: Department of Orthopedics, The First Hospital Affiliated to Dalian Medical University, Dalian, Liaoning 116011, P.R. China.
authors.forename: Decheng
authors.initials: D
authors.lastname: Lv
mesh_terms: D000818:Animals; D015153:Blotting, Western; D001854:Bone Marrow Cells / Q000378:metabolism*; D002454:Cell Differentiation; D002469:Cell Separation; D048430:Cell Shape; D048049:Extracellular Signal-Regulated MAP Kinases / Q000378:metabolism; D005260:Female; D016222:Fibroblast Growth Factor 2 / Q000235:genetics* / Q000378:metabolism; D005434:Flow Cytometry; D005822:Genetic Vectors / Q000378:metabolism; D005904:Glial Fibrillary Acidic Protein / Q000378:metabolism; D006801:Humans; D016130:Immunophenotyping; D016086:Lentivirus / Q000378:metabolism; D008297:Male; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000378:metabolism*; D020932:Nerve Growth Factor / Q000235:genetics* / Q000378:metabolism; D009474:Neurons / Q000166:cytology*; D010751:Phosphopyruvate Hydratase / Q000378:metabolism; D051057:Proto-Oncogene Proteins c-akt / Q000378:metabolism; D017207:Rats, Sprague-Dawley; D014161:Transduction, Genetic; D014162:Transfection; D014404:Tubulin / Q000378:metabolism
chemical_list: D005904:Glial Fibrillary Acidic Protein; C488416:Tubb3 protein, rat; D014404:Tubulin; D016222:Fibroblast Growth Factor 2; D020932:Nerve Growth Factor; D051057:Proto-Oncogene Proteins c-akt; D048049:Extracellular Signal-Regulated MAP Kinases; D010751:Phosphopyruvate Hydratase