Isolation, Induction of Neural and Glial Differentiation and Evaluating the Expression of Five Self Renewal Genes in Adult Mouse Neural Stem Cells

Purpose: Isolation, induction of neural and glial differentiation and evaluating the expression of Nucleostemin, ZFX, Hoxb-4, Sox-9 & Bmi-1 self renewal genes in adult mouse neural stem cells. Materials and Methods: Breifly, for isolation of neural stem cells, frontal part of adult mouse brain was minced in PBS and digested by enzyme solution, containing hyaloronidase and trypsin. Isolated cells were cultured in medium supplemented by EGF, bFGF. After one week, primary neurospheres were appeared in culture medium which differentiated into neural and neuroglial cells, after transferring to poly-L-ornyethin coated dishes. Differentiated cells were examined by morphological, immunocytochemical and molecular evaluations. Results: Our results showed that isolated cells from periventricular area of mouse adult brain can be expanded in medium containing EGF, bFGF and Fetal calf serum and differentiated into neural and neuroglial cells. Immunocytochemical studies for GFAP, MAP2 and O4 genes, proved the differentiation. Our results also showed that Nucleostemin, ZFX, Hoxb-4, Sox-9 & Bmi-1 are expressed in mouse neural stem cells. Conclusion: Mouse adult brain contains a type of stem cell which can be differentiated into neural and neuroglial cells. In addition, expression of Sox-9, Nucleostemin and Bmi-1 self renewal genes in undifferentiated neural stem cells can be considered as a proof for stem cell identity of isolated cells. This study is the first report which shows the expression of ZFX and Hoxb-4 genes in neural stem cells.