I've been using Methocult (I've tested both the so-called "complete methocult" and the "basal methocult") to grow BFU-E colonies from murine BM. I've tested several cell concentrations and like the Stem Cell Technologies protocol suggests, 0,6x10^5 cells in triplicate seems to be the best.
However, ever since those tests, I'm unable to get any BFU-Es and I've also realized that the current methocult lots are way more "waterish". I don't know if the problem is caused by this, but I was just wondering if there's any colleague out there who works successfully with murine BFU-Es on a regular basis and would kindly share with me any BFU-E secrets that might slip from my mind.
Thanks in advance.
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