Purify Astrocyte Culture from Neurons?

http://www.miltenyibiotec.com/en/products-and-services/macs-cell-separation/cell-separation-reagents/microbeads-and-isolation-kits/neural-cells/astrocyte-isolation-starter-kits.aspx

Yixiong Chen

Shanghai Jiao Tong University

Anuraj Nayarisseri

EMINENT BIOSCIENCES

Alfredo Pereira Junior

São Paulo State University

Nirmala Tilija Pun

Keimyung University

Joaquin M Fuster

University of California Los Angeles Medical School

Matthew L Neal

Florida International University

In my experience, neuronal cultures need a higher nutrient-rich media and using DMEM or MEM media without any neuronal supplements (e.g. B-27) will make it so that the neurons cannot survive in the culture. What media are you using for your mixed cultures?

Another method would be to use a separation kit, such as one offered by Miltenyi. They offer a GLAST-positive cell (astrocytes) isolation kit that would help purify your astrocyte cultures as well. (link provided below)

Hope this information helped.

1 Recommendation

Jordi Galiano-Landeira

Autonomous University of Barcelona

Hi Nesrin,

As it was very well pointed out by Matthew, if you want to get rid of the NPCs in your culture you have to use astrocyte culture media. It has DMEM, high glucose and 10% heat-inactivated fetal bovine serum. You have to feed them during some days allowing astrocytes to grow as well as neurons to die.

You can find a protocol very well described attached in the following reference:

Schildge, S., Bohrer, C., Beck, K., & Schachtrup, C. (2013). Isolation and culture of mouse cortical astrocytes. Journal of visualized experiments: JoVE, (71).

I hope you find it useful,

Jordi.

Mohammad Alqudah

Arabian Gulf University

see this video protocol in JOVE:

Article Isolation and Culture of Mouse Cortical Astrocytes

Check this reference for the protocol used to culture astroglia & microglia:

Article Regulation of Nerve Growth Factor Release by Nitric Oxide th...

I hope this can help.

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