title: Production of stable GFP-expressing neural cells from P19 embryonal carcinoma stem cells.
abstract: Murine P19 embryonal carcinoma (EC) cells are convenient to differentiate into all germ layer derivatives. One of the advantages of P19 cells is that the exogenous DNA can be easily inserted into them. Here, at the first part of this study, we generated stable GFP-expressing P19 cells (P19-GFP+). FACS and western-blot analysis confirmed stable expression of GFP in the cells. We previously demonstrated the efficient induction of neuronal differentiation from mouse ES and EC cells by application of a neuroprotective drug, selegiline In the second part of this study selegiline was used to induce differentiation of P19-GFP+into stable GFP-expressing neuron-like cells. Cresyl violet staining confirmed neuronal morphology of the differentiated cells. Furthermore, real-time PCR and immunoflourescence approved the expression of neuron specific markers. P19-GFP+cells were able to survive, migrate and integrated into host tissues when transplanted to developing chick embryo CNS. The obtained live GFP-expressing cells can be used as an abundant source of developmentally pluripotent material for transplantation studies, investigating the cellular and molecular aspects of early differentiation.
authors.affiliation: Cellular & Molecular Research Center, Shahrekord University of Medical Sciences, PO Box 8815713471, Shahrekord, Iran. Electronic address: shirzadeh@yahoo.com.
authors.forename: Hedayatollah
authors.initials: H
authors.lastname: Shirzad
authors.affiliation: Department of Biology, Faculty of Sciences, University of Isfahan, PO Box 8174673441, Isfahan, Iran. Electronic address: f.smaeili@sci.ui.ac.ir.
authors.forename: Fariba
authors.initials: F
authors.lastname: Esmaeili
authors.affiliation: Department of Anatomy, Tehran University of Medical Science, PO Box 1417653955, Tehran, Iran. Electronic address: sh-bakhshalizadeh@razi.tums.ac.ir.
authors.forename: Shabnam
authors.initials: S
authors.lastname: Bakhshalizadeh
authors.affiliation: Cellular & Molecular Research Center, Shahrekord University of Medical Sciences, PO Box 8815713471, Shahrekord, Iran. Electronic address: ebrahimi.marzeh@yahoo.com.
authors.forename: Marzieh
authors.initials: M
authors.lastname: Ebrahimie
authors.affiliation: School of Animal and Veterinary Sciences, The University of Adelaide, SA, Australia; School of Information Technology and Mathematical Sciences, Division of Information Technology, Engineering and the Environment, University of South Australia, Adelaide, Australia; Institute of Biotechnology, Shiraz University, Shiraz, Iran; School of Biological Sciences, Faculty of Science and Engineering, Flinders University, Adelaide, Australia; School of Medicine, The University of Adelaide, Adelaide, Australia. Electronic address: esmaeil.ebrahimie@adelaide.edu.au.
authors.forename: Esmaeil
authors.initials: E
authors.lastname: Ebrahimie
mesh_terms: D000818:Animals; D018929:Cell Culture Techniques / Q000379:methods*; D002454:Cell Differentiation / Q000187:drug effects; D002645:Chickens; D054278:Embryonal Carcinoma Stem Cells / Q000187:drug effects / Q000473:pathology* / Q000637:transplantation; D005453:Fluorescence; D005455:Fluorescent Antibody Technique; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects; D049452:Green Fluorescent Proteins / Q000378:metabolism*; D051379:Mice; D009474:Neurons / Q000187:drug effects / Q000378:metabolism* / Q000473:pathology*; D012642:Selegiline / Q000494:pharmacology; D014162:Transfection
chemical_list: D049452:Green Fluorescent Proteins; D012642:Selegiline
keywords: Antiparkinson drugs; GFP; Neuronal differentiation; Selegiline; Stem cells; Synaptophysin