Comparation of enhanced green fluorescent protein gene transfected and wild-type porcine neural stem cells.

The aim of this study was to transfect and express the enhanced green fluorescence protein (EGFP) gene into porcine neural stem cells (NSCs) to determine whether EGFP can be used as a marker to monitor NSCs. NSCs were isolated from embryonic day 30 fetal pig brain and transfected with EGFP gene using lipofection. Transfected and wild-type NSCs were induced to differentiate into cells of neuronal and myogenic lineages. Markers of passage three NSCs and their differentiated cells were tested by reverse transcription polymerase chain reaction. The results showed that EGFP could be expressed in NSCs and the differentiated cells. NSCs expressed Nestin, NogoA, DCX, Hes1, Oct4, CD-90 and Sox2. NSCs could differentiated into astrocyte (GFAP(+)), oligodendrocyte (GalC(+)), neuron (NF(+), NSE(+) and MAP2(+)) and myocyte (myf-6(+) and myoD(+)). We concluded that EGFP can be used as a marker in monitoring NSCs.

[1]  Mahendra Rao,et al.  SOX2, a Persistent Marker for Multipotential Neural Stem Cells Derived from Embryonic Stem Cells, the Embryo or the Adult , 2004, Developmental Neuroscience.

[2]  H. Wiesmann,et al.  Decreased CD90 expression in human mesenchymal stem cells by applying mechanical stimulation , 2006, Head & face medicine.

[3]  W. Hwang,et al.  Production of Nuclear Transfer-Derived Piglets Using Porcine Fetal Fibroblasts Transfected with the Enhanced Green Fluorescent Protein1 , 2003, Biology of reproduction.

[4]  R. Barker,et al.  The Potential for Circuit Reconstruction by Expanded Neural Precursor Cells Explored through Porcine Xenografts in a Rat Model of Parkinson's Disease , 2002, Experimental Neurology.

[5]  Jürgen Winkler,et al.  Transient expression of doublecortin during adult neurogenesis , 2003, The Journal of comparative neurology.

[6]  P. Halada,et al.  A proteomic approach to studying the differentiation of neural stem cells , 2007, Proteomics.

[7]  M. Chalfie,et al.  Green fluorescent protein as a marker for gene expression. , 1994, Science.

[8]  R. Prather,et al.  Multipotent adult progenitor cell lines originating from the peripheral blood of green fluorescent protein transgenic swine. , 2006, Stem cells and development.

[9]  I. Kirov,et al.  Expression of Neurodevelopmental Markers by Cultured Porcine Neural Precursor Cells , 2005, Stem cells.

[10]  J. Thomson,et al.  Embryonic stem cell lines derived from human blastocysts. , 1998, Science.

[11]  S. Antonini,et al.  Porcine embryonic stem cells: Facts, challenges and hopes. , 2007, Theriogenology.

[12]  T. Nagai,et al.  A Novel Method for the Production of Transgenic Cloned Pigs: Electroporation-Mediated Gene Transfer to Non-Cultured Cells and Subsequent Selection with Puromycin1 , 2005, Biology of reproduction.

[13]  Ning Li,et al.  Production of porcine cloned transgenic embryos expressing green fluorescent protein by somatic cell nuclear transfer , 2006, Science in China Series C: Life Sciences.

[14]  Teoan Kim,et al.  Expression of enhanced green fluorescent protein (EGFP) and neomycin resistant (NeoR) genes in porcine embryos following nuclear transfer with porcine fetal fibroblasts transfected by retrovirus vector , 2000, Molecular reproduction and development.