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[C.T]

I have been having some problems with my monocyte cell cultures and wondered if anyone had any ideas.

I separate PBMCs from human blood using histopaque, then use miltenyi CD14 beads to get the CD14+ cells.  After 48hr cell culture between 40-50% of my cells are dead.

I culture in RPMI supplemented with 10% FCS, L-glut and pen/strep, and have also tried adding 2-ME - with no benefit.

Any ideas to increase monoccyte cell survival?