The striatum or the V-SVZ was dissected from P4 to P7 WT mice and cultured as previously described (11 (link)). To increase the GFAP and Robo2 expression, immediately after changing the medium to FreeStyle (Thermo Fisher Scientific) containing penicillin/streptomycin (50 U/ml), the striatal astrocytes were incubated under 0 to 2% O2 (AnaeroPack Kenki, Mitsubishi Gas Chemical Company, Tokyo, Japan) for 20 hours and then cultured in Dulbecco’s modified Eagle’s medium containing 10% fetal bovine serum, penicillin/streptomycin (50 U/ml), and 2 mM l -glutamine for 0 to 4 days, depending on the experiment.
Protocol detail
Culturing Striatal Astrocytes with Hypoxia
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Astrocytes
Cultured medium
Eagle
Fetal bovine serum
Gfap
Glutamine
Mice
Penicillin
Streptomycin
Striatal
Corresponding Organization : Jichi Medical University
Other organizations : Ho Chi Minh City Medicine and Pharmacy University
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Variable analysis
independent variables
- Oxygen concentration (0 to 2% O2)
- GFAP expression
- Robo2 expression
- Culture medium (FreeStyle, Dulbecco's modified Eagle's medium with 10% fetal bovine serum)
- Penicillin/streptomycin concentration (50 U/ml)
- L-glutamine concentration (2 mM)
- Culture duration (0 to 4 days)
- Positive control: Not explicitly mentioned.
- Negative control: Not explicitly mentioned.
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