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These multipotent adherent progenitor cells (MultiStem®) are derived from mononuclear bone marrow cells that are depleted of CD45(+)/glycophorin-A (GlyA)(+) cells, using micromagnetic beads.
Cells are grown in CD45−GlyA− cell expansion medium supplemented with EGF (10 ng/mL) and PDGF-BB (10 ng/mL) and FBS (18%), on plates coated with fibronectin (10 ng/mL), laminin, or type IV collagen. Expansion medium: low-glucose DMEM (60%), MCDB-201 (40%), insulin (x1) transferrin selenium, linoleic acid, bovine serum albumin (x1), dexamethasone (10−9 M), ascorbic acid 2-phosphate (10−4 M), penicillin (100 U), and streptomycin (1000 U). Once adherent cells are more than 50% confluent, they are detached with trypsin-EDTA (0.25%) and are replated at a 1:4 dilution.