Neuronal Differentiation of mouse embryonic stem cells
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The major goal of this study is to differentiate mouse ESCs into neural progenitor cells (NPCs) and investigate the inducibility of a reporter transgene during this neuronal differentiation. I observed that this particular EGFP (enhanced green fluorescence protein) carrying ESC clone (C8) exhibited a very strong transgene inducibility comparing with the other EGFP clones and also this cell clone retained its transgene inducibility upon neural differentiation: 98% of the drug-induced cells were GFP positive after 6 day of differentiation. In addition, I observed that these ESC derived differentiated cells were also able to form neuronal networks and they expressed neural specific genes (Pax6 and Neurog2).
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BMP Bone Morphogenetic Proteins , bFGF Basic Fibroblast Growth Factor , CNS Central Nervous System , DOX Doxycycline , DMEM Dulbecco's Modified Eagle Medium , DMSO Dimethyl Sulfoxide , EGFP Green Fluorescence Protein , ESCs Embryonic Stem cells , FACS Fluorescence Activated Cell Sorting , FBS Fetal Bovine Serum , FGF Fibroblast Growth Factor , GFAP Glial Fibrillary Acidic Protein , G418 Genetic , HSCs Hematopoietic Stem Cells , hESCs Human Embryonic Stem Cells , ICM Inner Cell Mass , iPSCs Induced Pluripotent Stem Cells , IKMC International Knockout Mouse Consortium , KSR Knockout Serum Replacement , LIF Leukemia Inhibitory Factor , mESCs Mouse Embryonic Stem Cells , MEF Mouse Embryonic Fibroblast , N2B27 Neurobasal plus B27 supplement , NPCs Neuro Progenitor Cells , PBS Phosphate Buffered Saline , Q- PCR Quantitative Polymerase Chain Reaction , RGPs Radial Glial Progenitor , TA Transactivator