Hi. I used mixed glial culture, then shake the primary microglia off and replate them in 96 well plate for functional assay. I saw different morphology between WT cell and gene A knockout cell. I enclosed here the CD11b staining picture of both cell types. The WT cell more look like the ramified shape, and the gene A KO cell looks more like an amoeba shape. What is the normal morphology of primary microglia from mixed glial culture? Does the gene A KO cell more like activated microglia?
Thanks!
Hi, I have the same problem as Staffan has just mentioned. Nevertheless, I attach a photo of microglia two days after being shaken; they are wt and unstimulated. To my experience, cell morphology could be changed after stimulation (e.g. by LPS) so the same could be possible after some gene manipulation.
Update: I've just succeed to download your pictures. At first glance, your KO microglia seem to have different morphology. In particular, round flattened cells seem to be more frequent. I usually see this "amoeboid" morphotype in my cells after some pro-inflammatory stimulation connected to M1 microglia polarization, see reference attached.
Article Bipolar/rod-shaped microglia are proliferating microglia wit...
Hi, is it possible to have new pictures or relod them because as the others I can't open them. Really difficult to see something .....
Thank you all for answering.
To Tony and Staffan, I re-upload the pictures. Hope this time could work.
Yes, it's better like that.
Based on our previous works and on what we are currently doing, in control conditions, microglial cells look like a ramified shape (as in your WT condition). After pro-inflammatory stimulations, microglial cells appear to change of morphology, to modify phagocytosis capacities and to express/release numerous mediators. In attachment, you will find one of our paper on glial cultures.
Concerning your Gene A KO microglial cells, they look like activated. It will be simple to compare and confirm the two types of microglial cells status (in term of activation) by performing ICC analyses (F4/80 will be higher in activated cells) and/or by real-time PCR (evaluation of Iba1 or CD11b expression as well as pro-inflammatory gene expressions like Nos2, Tnf, Cxcl10, Ptgs2, ...).
Xianyuan Xiang Have you solve this problem? I have met the same phenomenon, however, there is no paper telling me what it is. Have you identified this cell type? is it really microglia, or macrophage or others? Wish for your reply!
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