Cell and Organ Transplantology. 2014; 2(2):155-157.
A protocol for isolation of fetal neural progenitor cells from mouse hippocampus
Tsupykov О. М.
Bogomoletz Institute of Physiology NAS Ukraine, Кyiv, Ukraine
State Institute of Genetic and Regenerative Medicine NAMS Ukraine, Kyiv, Ukraine
Culture of neural stem/progenitor cells are widely used to study the characteristics of these cells under controlled conditions in vitro as well as to study the cellular and molecular mechanisms of CNS diseases and develop strategies for their treatment.
This paper provides a detailed protocol to isolate of fetal (E17-18) neural progenitor cells (NPCs) of mouse hippocampus. The technique is based on the use of centrifugation of hippocampal cells suspension in Percoll density gradient to obtain purified NPCs fractions. The cells are cultured in serum-free medium in a monolayer, which creates conditions for more equitable access of FGF-2 to the cells. This method provides a homogeneous population of undifferentiated progenitors from fetal mouse hippocampus.
|1. Ma DK, Bonaguidi MA, Ming GL, et al .Adult neural stem cells in the mammalian central nervous system. Cell Res. 2009; 19(6):672-682.
|2. Ming GL, Song H. Adult neurogenesis in the mammalian brain: significant answers and significant questions. Neuron. 2011; 70(4):687–702.
|3. Blakemore WF, Franklin RJ. Transplantation options for therapeutic central nervous system remyelination. Cell Transplant. 2000; 9(2):289–294.
|4. Reynolds BA, Tetzlaff W, Weiss S. A multipotent EGF-responsive striatal embryonic progenitor cell produces neurons and astrocytes. Neurosci. 1992; 12:4565–4574.|
|5. Kukekov VG, Laywell ED, Suslov O, et al. Multipotent stem/progenitor cells with similar properties arise from two neurogenic regions of adult human brain. Exp. Neurol. 1999; 156(2):333-344.
|6. Pagano SF, Impagnatiello F, Girelli M, et al. Isolation and characterization of neural stem cells from the adult human olfactory bulb. Stem Cells. 2000; 18(4):295-300.
|7. Vescovi AL, Snyder EY. Establishment and properties of neural stem cell clones: plasticity in vitro and in vivo. Brain Pathol. 1999; 9(3):569-598.
|8. Palmer TD, Ray J, Gage FH. FGF-2-responsive neuronal progenitors reside in proliferative and quiescent regions of the adult rodent brain. Mol. Cell Neurosci. 1995; 6:474–486.
|9. Jenny B, Kanemitsu M, Tsupykov O, et al. Fibroblast growth factor-2 overexpression in transplanted neural progenitors promotes perivascular cluster formation with a neurogenic potential. Stem Cells. 2009; 7:1309–1317.
|10. Babu H, Claasen J H, Kannan S, et al. A protocol for isolation and enriched monolayer cultivation of neural precursor cells from mouse dentate gyrus. Front. Neurosci. 2011; 5: article 89.
|11. Kempermann G, Jessberger S, Steiner B, et al. Milestones of neuronal development in the adult hippocampus. Trends Neurosci. 2004; 27:447–452.
Tsupykov ОМ. A protocol for isolation of fetal neural progenitor cells from mouse hippocampus. Cell and Organ Transplantology. 2014; 2(2):155-157. doi: 10.22494/COT.V2I2.32