What is the best protocol to culture/expand mesenchymal stem cell in the lab?
Mesenchymal stem cells (MSCs) are non-hematopoietic cells capable of self-renewal and differentiation into bone, cartilage, muscle and related cells. Similar to hematopoetic stem cells, MSCs are rare (0.001% of bone marrow cells). Several studies have been carried out to optimize culture and expansion conditions for MSCs. Commercial suppliers now make optimized media for human and mouse MSC culture. Fetal calf serum used in normal cell culture is NOT recommended for stem cells. Autologous serum has been used to expand MSCs for clinical purposes, but serum-free media is desirable. The process involves: 1) flushing out all cells from the bone marrow; 2) removing mature cells of different lineage using antibodies; 3) Using density gradient centrifugation to isolate remaining live stem cells; 4) In-vitro expansion of MSCs using optimized media; and 5) Phenotyping MSCs for purity.
