Mesenchymal stromal cells (MSCs) are routinely harvested using detachment reagents such as trypsin-EDTA, TypLE Express Enzym, TrypZean, or collagenase. Detachment reagent concentration and detachment time must be carefully optimized for each culture system to enable maximum recovery of cells while preserving high cell quality. A combination of different detachment reagents and the integration of hydrodynamic forces can facilitate the detachment of the cells from each other as well as from the microcarrier. Note, if cells are growing confluent on the microcarrier it might take longer to detach them, which ultimately could affect their quality.