Any advice for Astrocytic markers for FACS?

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Ian Clift

Astrocytes act as Antigen Presenting Cells (APCs) which means that using several readily available immunologic markers might work.

Sun Yung

CD44 expression identifies astrocyte-restricted precursor cells, so we use MACS CD44+ beads to enrich our Astrocyte Progenitor population first prior to differentiating into astrocytes.

You may want to check out this too:

https://www.google.co.uk/url?sa=t&rct=j&q=&esrc=s&source=web&cd=1&cad=rja&uact=8&ved=0ahUKEwjPh42X77zJAhUExxQKHT_aBl4QFgguMAA&url=https%3A%2F%2Fwww.miltenyibiotec.com%2F~%2Fmedia%2FFiles%2FNavigation%2FResearch%2FNeuroscience%2FWPF_customer_report_Jungblut.ashx&usg=AFQjCNEqSFmgjlSI13PNVxbdzPvIFWf2zw&sig2=2vZC754J6TNXrS25iqWysA

Lech Kaczmarczyk

I would consider Gja1 (Cx43).

Oksana Tuchina

Hi Amit! GFAP and S100 beta are intracellular markers, while my aim is to specifically stain live cells. Sun and Lech, thank you for your suggestions! I havent thought about connexins.

Li Li

Hi Oksana,

How is your experience with GLAST sorting of astrocytes? I tried CD44-APC (from eBioscience), but this didn't work well because of strong background.

Hi Li! I think GLAST works well (we use the one from miltenyibiotec) in order to sort live astrocytes from cell suspension of a 3-4 day rat brain. Strong background in APC channel? Maybe there was a lot of unspecific staining? I usually get high background in FL1 (green channel), and that is especially when working with fixed cells.

Thanks for the answer! My CD44 does not give two populations. Maybe my cells (iPSC derived) are too mature and CD44 might work better for progenitor cells. I'll try GLAST instead. Good luck with your sorting.