Five days ago, I harvested splenocytes from C57BL/6 mice by 70 um cell strainer and in order to remove any adherent cells including macrophages, I cultured them in a treated 25 flask for 24 hours (by RPMI 1640). The next day I transferred the suspended cells into 24 well plates. Today, I harvested the cells and media, at the following, I found some fibroblast-like spindle shaped cells attached to the surface of the wells.
What are these cells? and why they have not removed at the first day? May it be due to using RPMI 1640 cell culture medium?
(The following enclosed files are amateur photos from the cells taken by my cell phone camera).
They look like macrophages, that we get after differentiation of monocytes in vitro. You could maybe have had some monocytes left in the cell suspension, when you removed it from the flask and these may have differentiated into macrophages during the longer culture in the 24 well plate.
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