Protocol for the Differentiation of Mouse Neural Stem Cells into Astrocytes

Astrocytes are a type of glial cell found in the central nervous system (CNS), including the brain and spinal cord. They constitute the largest population of glial cells and play essential roles in the development, maintenance, and functioning of the CNS. It is important to establish effective methods to differentiate reliable stem cells, such as neural stem cells, into mature astrocytes.

• A 12-mm cell culture slide is placed in each well of a 4-well plate, and 500 μL of PDL at 100ug/mL is added to each well and left to dry at room temperature overnight.
• 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 in four-well plates in which laminin is discarded, and the cell density is 1×10⁴ cells/ml, at which time the culture medium is Astrocyte M.
• Astrocyte M is replaced in half quantities every 2 days by day 7 when differentiation is complete and astrocytes can be observed.

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• Astrocyte differentiation medium is stable for 4 weeks when stored at 2-8°C in the dark.
• Replace the used medium every 3-4 days.

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