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Primary neuron culture can be easily prepared.
Neuron Culture

The concept of the product family is to make preparation of rat or murine primary neuron culture easy for anyone , which
is generally thought as difficult. Technical guidance was given by Sumitomo Bakelite Co., Ltd. for commercialization.
This is a subsequent product of our neuron culture medium, and neuron dispersion solution series.

1. Link

2-1. Neuron Culture Medium

This medium is for nerve cells and enables stable cultivation of primary nerve cell. It is optimized for central nervous
system cell culture. This product contains a culture supernatant of rat glial cell.

2-2. Features

■Rapid neurite outgrowth

Neuron Culture Medium: for 14 days
A conventional medium: for approx. 1 month

 

 x about 1/2 of the conventional culture period
■Low density culture

Neuron Culture Medium: 0.1 x 106 cells/mL
A conventional medium: 0.5 to 1.0 x 106 cells/mL 

 

 x 1/5 to 1/10 of the conventional cell number
■Ready to Use
Culture is readily feasible with medium only. No additional supplements are necessary.

2-3. Confirmation of neurite outgrowth (MAP2 immunostaining)

 

Faster neurite outgrowth was confirmed in Neuron Culture Medium (Wako) than in a competitor’s medium.


Experimental conditions
Cell number 0.1 x 106 cells/mL (Dispersed from hippocampus of a fetal mouse (embryonic day 18.5)
Culture scale 500 μL/well (Polylysine coated glass bottom dish)
Culture condition A half of the medium is changed on Day 3 and Day 7 (without addition of Ara-C)

Data provided by
Dr. Hirotaka James Okano and Mr. Yuki Ogawa, Division of Regenerative Medicine, Jikei University School of Medicine (Japan)

2-4. Confirmation of neurite outgrowth (MAP2 immunostaining)

 

Better neurite outgrowth and a higher cell number was confirmed in neurons cultured with the product
than those with a competitor’s medium.

Experimental conditions : Same as above.
Data: Same as above.

2-5. Evaluation of maturity of the neurons: Confirmation of the neurite spines

Dendritic spines are observed in mature neurons cultured by Neuron Culture Medium.
→Mature time of Neuron Culture Medium is shorter than a competitor’s medium.

Experimental conditions : Same as above.
Data: Same as above.

2-6. Evaluation of maturity of neurons: Confirmation of the axons and neurites

MAP2:A marker of neurites; AnkyrinG:A marker of the proximal part of the neuroaxis

Normal cell maturation was confirmed based on several neurites and an axon extended from the
cell body of the neurons cultured with the product.

 

Experimental conditions : Same as above.
Data: Same as above.

2-7. Product List

Product Name Pkg. Size Grade Wako Cat. No. Storage Condition
Neuron Culture Medium 100 mL for Cell Culture

148-09671

Keep at -80℃

3-1. Neuron Dissociation Solutions

Dispersion and isolation of neurons from rat and mouse central nervous tissues are feasible with the reagents. It consists of 3 solutions (Enzyme Solution, Dispersion Solution, Isolation Solution). By using the product, the reagent preparation time can be saved and neurons can be easily isolated with high survival rate.

3-2. Features

■Neurons can be easily and steadily isolated

■Ready to Use

3-3. Protocol

1 Slice the brain tissues into 1 to 2 mm fragments and put them into a 15 mL centrifuge tube.
 
2 Add Enzyme Solution and incubate at 37°C for 30 minutes.
 
3 Pipette in the enzyme solution and disperse the tissue.
 
4 Centrifuge at 900 to 1,000 rpm for 4 minutes.
 
5 Remove the supernatant and add Dispersion Solution to disperse.
 
6 Gently add Isolation Solution beneath the dispersion solution.
Prepare the 2-layer solution with the dispersion solution (upper layer)
and isolation solution (lower layer) (See the right figure).
 
7 Centrifuge at 800 to 900 rpm for 5 minutes.
 
8 After removing the supernatant, disperse the tissues with Neuron Culture Medium (Wako Cat. No.148-09671) again and culture them on a polylysine coated culture dish.
When growth of glial cells is abundant, add Ara-C to them. (Day 3-5: 5~10 μmol/L).

 

3-4. Product Lists

 

Product Name Pkg. Size Kit contents Grade Wako Cat. No. Storage Condition
Neuron Dissociation Solutions 4 sets

Enzyme Solution

Dispersion Solution

Isolation Solution

5.0mL x 4

5.0mL x 4

5.0mL x 4

for
Cell Culture
291-78001 Keep at -80°C
Neuron Dissociation Solutions S 10 sets

Enzyme Solution

Dispersion Solution

Isolation Solution

2.5mL x 10

2.5mL x 10

2.5mL x 10

297-78101

3-5. Related Product Lists

Product Name Pkg. Size Grade Wako Cat. No. Storage Condition
Cytosine-1-β-D(+)-arabinofuranoside
(Synonyms:Ara-C, Cytarabine)

100 mg

500 mg

1 g

for
Biochemistry

030-11951

034-11954

036-11953

Keep at 2-10°C
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