title: Unrestricted somatic stem cells as vehicle for nerve growth factor gene transfer.
abstract: OBJECTIVE
  Nerve growth factor (NGF), a member of the neurotrophic factor family, plays a critical role in the maintenance and regeneration of different types of neurons. To overcome drastic challenges in the peripheral delivery of NGF, transplantation of NGF secreting stem cells to the target site of an injury may be an effective procedure. Unrestricted somatic stem cells (USSCs), a subtype of umbilical cord blood (UCB) stem cells, have shown promise for gene therapy purposes, and proper results have been observed from transplantation experiments in neurodegenerative disorders. Based on the considerable potential of USSCs for gene delivery applications, the goal of the current study was to establish a betaNGF gene containing USSCs, which is able to secrete functional recombinant betaNGF protein.
  
  
  METHODS
  Unrestricted somatic stem cells were isolated from UCB and were cultured in a DMEM medium. The betaNGF gene was cloned in the EFalpha-promoting lentiviral vector, and virus production was performed as a third generation lentivirus packaging system. Enzyme-linked immunosorbent assay, real-time PCR, co-culture, MTT, and immunocytochemistry assays were performed to evaluate the genetically engineered USSCs.
  
  
  RESULTS
  Overexpression of betaNGF gene in human USSCs created a USSC line that is able to secrete high amounts of functional betaNGF protein. betaNGF-secreting USSCs showed a high rate of viability along with acceptable immunological and morphological properties for transplantation into the nervous system.
  
  
  CONCLUSIONS
  Long-term expression of functional betaNGF, high viability of betaNGF producing USSCs, and expression of primary neuronal markers suggest that USSCs may be useful to deliver betaNGF into targeted sites of the nervous system in neurodegenerative disorders.
authors.affiliation: Department of Developmental Biology, Faculty of Biological Science, Kharazmi University, Tehran, Iran. soleim_m@modares.ac.ir
authors.forename: Hanieh
authors.initials: H
authors.lastname: Jalali
authors.forename: Kazem
authors.initials: K
authors.lastname: Parivar
authors.forename: Mohammad
authors.initials: M
authors.lastname: Nabiuni
authors.forename: Masoud
authors.initials: M
authors.lastname: Soleimani
mesh_terms: D053687:Adult Stem Cells / Q000378:metabolism*; D000818:Animals; D002470:Cell Survival / Q000502:physiology; D018920:Coculture Techniques; D005312:Fetal Blood / Q000166:cytology* / Q000378:metabolism*; D018014:Gene Transfer Techniques; D057809:HEK293 Cells; D006801:Humans; D020932:Nerve Growth Factor / Q000096:biosynthesis / Q000235:genetics*; D016716:PC12 Cells; D051381:Rats
chemical_list: D020932:Nerve Growth Factor