Biology Reference
In-Depth Information
Ultra-glutamine (2 mM) and antibiotics to a final concentra-
tion of 100 U/ml Penicillin and 100 μg/ml Streptomycin;
Packaging plasmids and transfer vector;
Tris-EDTA (TE) solution.
Tris (10 mM, pH 8.0) diluted 1:10 with H 2 0, 0.22 μm filtered
and stored at 4 °C.
EDTA (0.5 M, pH 8.0).
The TE solution is subsequently diluted 2:1 in H 2 0;
2× Hydroxyethyl piperazineethanesulfonic acid (HEPES)—buffered
saline (HBS)
NaCl (281 mM),
HEPES (100 mM),
1.5 mM Na 2 HPO 4 [pH 7.12] 0.22 μm filtered and stored at
−20 °C;
CaCl 2 solution 2.5 M: 0.22 μm filtered and stored at −20 °C;
PBS 1X;
Trypsin;
Hexadimethrine bromide (Polybrene).
Lentiviral particles;
2.3 NPC
Transduction by
Lentiviral Infection
In Vitro
37 °C, 5 % CO 2 humidified incubator;
Hood (Cat2);
NPC growth medium
NeuroCult ® Proliferation Kit (mouse; Stem cell technologies)
supplemented with
Mouse recombinant epidermal growth factor (EGF, 20 ng/ml);
Mouse recombinant fibroblast growth factor (FGF)-2, 10 ng/ml;
Heparin at a final concentration of 0.0002 %; w/v; equals
2 μg/ml and
Antibiotics to a final concentration of 100 U/ml Penicillin and
100 μg/ml Streptomycin;
Accumax.
Heat lamp;
Tailveiner Restrainer;
1 cc or 1/2 cc Insulin Syringes (Terumo).
2.4 Transplantation
of Transduced NPCs
2.4.1 NPC
Transplantation Through
i.v. Injection
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