Biology Reference
In-Depth Information
4. Glucose-enriched medium: D-MEM with 4.5 g/L glucose
(Gibco).
5. Neurobasal medium (Gibco).
6. Gentamicin solution: 50 mg/mL gentamicin sulfate solution
(Gibco).
7. B-27 supplement without antioxidants (minus AO, Gibco).
8. Epidermal growth factor (EGF): 20
g/mL recombinant
human EGF (Preprotech), dissolved in water and stored at
−20 °C.
9. Fibroblast growth factor (FGF): 20
μ
g/mL recombinant
human FGF (Preprotech) dissolved in 5 mM Tris, pH 7.6 and
stored at −20 °C.
10. Alpha-tocopherol: 1 mg/mL alpha-tocopherol (SIGMA) dis-
solved in ethanol and stored at −20 °C.
11. Alpha-tocopherol-acetate: 1 mg/mL alpha-tocopherol-acetate
(SIGMA) dissolved in ethanol and stored at −20 °C.
12. GlutaMAX solution: 200 mM glutaMAX.
13. Forskolin: 10 mM forskolin (SIGMA) dissolved in ethanol.
14. N6,2
μ
-cyclic monophosphate
sodium salt (db-cyclic AMP): 1 M db-cyclic AMP (SIGMA)
dissolved in water.
15. poly-
L
-ornithine: 0.01 % poly-
L
-ornithine solution (SIGMA).
16. Fibronectin: 1 mg/mL human plasma fi bronectin (Chemicon,
Millipore).
17. T-25 cm
2
fl asks with ventilated cap.
18. T-75 cm
2
fl asks with ventilate cap.
19. Cell culture dishes: 60 and 100 mm dishes.
20. Accutase solution (SIGMA) stored at −20 °C.
21. Penicillin/streptomycin: 5,000 U/mL penicillin and
5,000
′
-
O
-Dibutyryladenosine 3
′
,5
′
g/mL streptomycin to be used 1:100.
22. Phosphate-buffered saline (PBS).
23. Expansion medium: 50 % glucose enriched medium, 50 % F-12
Ham, 2 % B-27 minus AO, 20 ng/mL EGF, 20 ng/mL FGF,
10
μ
μ
g/mL gentamicin, 1
μ
g/mL alpha-tocopherol, 1
μ
g/mL
alpha-tocopherol-acetate.
24. Dopaminergic differentiation medium: neurobasal medium
supplemented with 2 % B-27 minus AO, 10
μ
g/mL gentamicin,
2 mM glutaMAX, 100
μ
M db-cyclic AMP, 10
μ
M forskolin.
1. DNase I: 1 U/
μ
L DNase I amplifi cation grade (Life
2.2 Titration of HD
Adenoviruses by
Real-Time PCR
Technologies).
2. DNase I reaction buffer: 200 mM Tris-HCl, pH 8.4, 20 mM
MgCl2, 500 mM KCl, to be used 1:10.
