Usha Ks use a Tissue culture treated plate to plate your cells, it supports the adhesion of the cells and you will see more of attached cells that will differentiate into macrophages upon stimulation by M-CSF.

Hello Usha Ks,

Non-tissue culture treated plates are typically not designed for cell adhesion, hence you will observe lack of cell attachment.

You should use tissue culture-treated plates as they have a modified surface that promotes cell adhesion through electrostatic interactions. Using these plates is the easiest way to ensure monocyte attachment. 

If tissue culture-treated plates are not available, you can coat non-treated plates with ECM proteins like fibronectin or collagen. These proteins provide a natural substrate for cell attachment.

Once monocytes are adhered and viable, they can be polarized into M1 and M2 macrophages using specific cytokine cocktails. 

Best,

Usha Ks ,

When culturing CD14+ monocytes derived from human peripheral blood mononuclear cells (hPBMC), it is recommended to use tissue culture treated (TC treated) culture dishes, because TC treated culture dishes are hydrophilic and have positively charged groups on the surface, which is conducive to the attachment of monocytes and differentiation into macrophages.

The answer to this question comes from MedChemExpress (MCE) Technical Support.

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Hi everyone, Thanks for your answers.

I did try to culture CD14+ monocytes from hPBMCs on tissue culture-treated plates this time. I do see only a couple of cells adhere to the plate, but not all of them (pic attached- taken on 5th day of MCSF incubation). I still do not know what is wrong.

I bought these cells from StemCell Technologies (frozen). Cultured them in RPMI media with 10% FBS and 50ng/ml MCSF or GMCSF for 5 days.