Hello everyone. I cultured isolated PBMCs in Erythroid Expansion Medium for two weeks and then performed electroporation in my expanded PBMCs in order to reprogram them. The first 24h after the electroporation I cultured my cells in Erythroid Expansion Medium, then I changed the medium to DMEM/FBS and plated them in Matrigel coated wells. Now I have switched to E8 Medium+NaB, still culturing them in Matrigel coated wells.
Everything was normal, until I switched to DMEM/FBS and to coated wells. Immediately when I made the switch I started noticing these weird clumps amongst my transfected PBMCs. They do not seem to halter the differentiation process nor the viability of my cells. At first I thought that they were Matrigel clumps, but now I wonder whether they might be clumps of white blood cells. Have you ever seen something similar to this?
Thank you in advance for your time.
Hi Stella. It looks to me like it's just debris. Either from the matrigel or the FBS. I don't think it will hurt your cells at all.
Based on my limited experience in culturing suspending cells, mostly of lymphoma origin, the clumping appearance may reflect a vigourous viability; the behaviour corresponds with bottom attachment of monolayers.
Thank you everyone for your input. After changing the medium two times, the majority of these clumps is gone and my cells are still proliferating and differentiating as expected :)
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