I am now culturing the Bone marrow mononuclear cells (BMMC) in RPMI supplemented with 10% FBS, 2mM L-glutamine & PHA-M (as mitogen).

does anyone know:

1) how to subculture & cryopreseve the cell?

2) how long will it takes usually for the cells to confluent after cryopreserved & revive back?

3) what are the morphologies of this cell?

4) at which passage number should i stop using them? as mine at passage number 2 mostly they grow into mesenchymal type of cells.

5) i would like to compare the treatment between myeloid leukemia cells and normal bone marrow cells, so which type of cell type is the best for my study?

thank you very2 much for your time.