For research on anti-oxidization, anti-inflammation, analysis of cytokines and gliosis.

Astrocyte Culture kit (Rat)


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Background

Heterogeneous Role of glial cells : Neurotoxicity or Neuroprotection

Astrocytes are one of the glial cells in the brain and spinal cord. They are the most abundant cells in the brain. They perform many functions, including construction of the central nervous system, regulation of homeostasis of neurons and formation of the blood-brain barrier. 

It has been known that inflammation, hepatic failure, neurodegeneration leads to Alzheimer’s Type II Astrocyte and morphological changes of fibrous astrocytes, and eventually gliosis, causing damage to the central nervous system (CNS). 

In addition , it has been revealed that Astrocytes become neurotoxic by the stimulation of cytokines such as tumor necrosis factor (TNF-α), interleukin and interferon (IL) -1β of (IFN) γ , reactive oxygen species (ROS) and arachidonic acid metabolites and nitric oxide (NO) production1.

On the other hand, it has been shown that Astrocytes also have a neuroprotective role; as an antioxidant for Glutathione (GSH) in Parkinson’s disease, and inhibition of synaptic transmission due to the release of neurotransmitters such as Glutamic Acid, ATP and D-Serine2.

 

 

1j Blasko, I., Sta mpfer-Kountchev, M., Ro batscher, P., Ve erhuis, R., Eikelenboom, P., & Grubeck-Loebenstein, B. (2004). Aging cell, 3(4), 169-76.
2j Maragakis, N. J., & Rothstein, J. D. (2006). Nature clinical practice Neurology, 2(12), 679-89.
3j Sawada, M. (2009). Parkinsonism & related disorders, 15 Suppl 1, S39 41.
4j Johansson, C., Momma, S., & Clarke, D. (1999). Cell, 96, 25-34.

Application

The Astrocyte Culture kit is a kit which consists of formulated cryopreserved cells and culture medium optimized for culturing astrocytes. It can be used for anti-oxidization, anti-inflammation, analysis of cytokines and gliosis.

Kit component

• Astrocytes, Rat (embryos(E18-20))     1 x 106 cells/vial 
• Astrocyte Culture Medium                 250ml 
   (Cat. Number: PMC-ASTM-COS)

Components of Medium
ASTM is a complete medium designed for optimal culture for rat astrocytes in vitro. It is a sterile, liquid basal medium (D-MEM/F12) which contain essential and non-essential amino acids, vitamins, other organic compounds, trace minerals, inorganic salts, growth factors, hormones, fetal bovine serum, and antibiotics.

Materials required but not provided
• Various volume pipettes
• Culture dishes, plates or flasks

Protocol

1. Thaw the vial of cryoserved Astrocytes in a 37°C water bath for 2 minutes. Caution: Do not vortex the cells

2. Transfer thawed cells into a 50 ml centrifuge tube containing 10 ml of Astrocyte Culture Medium.

3. Rinse the vial with 1ml of cell suspension.

4. Centrifuge for 5 minutes at 4C at 200 x g.

5. After removing the supernatant, resuspend cells with 10ml volumes of medium.

6. Centrifuge for 5 minutes at 4C at 200 x g.

7. After removing the supernatant, resuspend cells in appropriate volume of medium.

8. Transfer the cell suspension to appropriate flasks, petri dishes or well plates. The recommended cells density is 0.5~1 x 104cells/cm2.

9. Incubate the plate at 37°C under 5% CO2, 100% humidity.

10. Every 2-3 days, replace the supernatant with warmed-up, fresh medium of cell culture and return cells to the incubator.

*After 3~5 days, astrocytes reach approximately 80~100% confluency.
*Astrocytes have weak adhesion to glass. When culturing on glass slides or glass dishes, the recommended cells density is 2~5 x 104cells/cm2. Astrocytes reach approximately 80~100% confluency in about one week.

Quality Control

The Astrocytes are characterized for each lot by Immunofluorescence of Glial Fibrillary Acidic Protein (GFAP)

Fig.1 Immunofluorescence of astrocytes A:GFAP, B:nuclear staining, C:bright field

References

> Miller, R. H., Ffrench-Constant, C., and Raff, M. C. (1989). The macroglial cells of the rat optic nerve. Annu. Rev. Neurosci. 12, 517-534. (2)

Product List

Precautions
• Please read the instructions carefully before beginning the culture.
• This kit is for research use only, not for human or diagnostic use.


Product Name Cat# Quantity Price

Astrocyte Culture kit DataSheet ,

PMC-AST01-COS

1 SET ¥119,600
$1595

To be used for research only. DO NOT use for human gene therapy or clinical diagnosis.


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