Protocol for the Differentiation of Mouse Neural Stem Cells into Neurons
GUIDELINE
The generation of the different neural lines originates in adult neural stem cells that can self-renew or differentiate into astrocytes, oligodendrocytes, or neurons in response to specific stimuli. The abilities of stem cells to self-renew and form different mature cells expand the possibilities of applications in cell-based therapies such as tissue re-composition in regenerative medicine, drug screening, and treatment of neurodegenerative diseases.
METHODS
- Place a 12-mm cell culture slide in each well of a 4-well plate, add 100 μg/mL of PDL 500 μL to each well, and leave overnight at room temperature.
- The PDL is aspirated, and the slides are rinsed with sterile water and left to dry at room temperature. 10 μg/mL of laminin 500 μL is added to each well, and the slides are allowed to stand overnight at 37°C and 5% CO2.
- Mouse neural stem cells are digested with 0.05% trypsin for 2 min and terminated with an equal volume of 10% FBS. Centrifugation is performed at 1000 rpm for 5 min, the supernatant is discarded and the cells are resuspended with Neuron M. The cells are inoculated at a cell density of 1×104 cells/ml in a four-well plate in which laminin is discarded.
- Neuron M is replaced in half quantities every 2 days, and by day 21, differentiation is complete, and neuronal cells with neurofilaments can be observed microscopically.
Creative Bioarray Relevant Recommendations
- Creative Bioarray provides our customers with a large and unique collection of high-purity, low-passage human and animal neural stem cells, including but not limited to Strain C57BL/6 Mouse Neural Stem Cells, Sprague-Dawley (SD) Rat Neural Stem Cells, Human Neural Stem Cells-cortex region, Human iPSC-Derived Neural Stem Cells, Human Neural Stem Cells-ventral mesencephalon region, Mouse Spinal Cord Neural Stem Cells, Rat Hippocampal Neural Stem Cells, and others.
NOTES
Do not expose cells to air at any time after they have differentiated into neurons.
