I isolate Monocytes and NK cells from blood and synovial fluid, using MACS method.
every time i got synovial fluid sample, i got a huge amount of cells in the sample, which deplete alot of my microbeads, and in the end i got bad purity!
I tried Ficoll gradient separation as primary step to get the mononuclear cells, before doing MACS (as a protocol that i found ), but it didn't work well as in blood!
And also, does any one find a difference between Microbeads suspension product and Lyophilized product , because since i ordered the second, i did not get one nice purity result!! it could be another reason, but it will be nice to share your expertise in this field ..
Dimah
Hi Dimah,
Can you please share the protocol for Macrophage culture from Synovial fluid & Synovial tissue??
Hi Anuj Kumar Pandey,
Actually, it was very difficult to isolate Monocytes from synovial fluid, because of the big abundance of neutrophils and another inflammatory cells (that we already isolate synovial fluid only when the synovium is inflammed). Even when i did the first step of f´Ficoll mono-nuclear cells Separation (it worked only when i used 15 ml Falcons not 50 ml Falcons).
My collegue has isolated the monocytes from synovial tissue by MACS method, with very low outcomes number.
i have depended on blood to get the monocytes , and i used Monocytes Isolation kit from Miltenyi Biotec.
Good luck :)
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