Designing clinical-grade integrated strategies for the downstream processing of human mesenchymal stem cells

Filtration methodologies for the clarification and concentration of human mesenchymal stem cells

“…As previously reported by our group [16], pore size is critical for the efficiency of the clarification step. High pore size filters (4 75 μm) permit higher cell recoveries, while lower pore size filters (30 μm) are subjected to microcarrier cake formation, entrapping part of the cell suspension, as previously observed by Nienow et al [10].…”

“…hMSC were routinely cultured in MesenCult s -XF Medium (STEMCELL ™ Technologies) supplemented with 2 mM L-Glutamine and propagated in tissue culture flasks (Thermo Scientific ™ Nunc ™ , Massachusetts USA), previously coated with MesenCult ™ -SF Attachment Substrate (STEMCELL ™ Technologies), at 37 1C in a humidified atmosphere of 5% CO 2 in air, as previously described [16]. At 70-80% cell confluency, the medium was removed and cells were washed with Dulbecco's phosphate-buffered saline (DPBS) and incubated with TrypLE Select (1X) dissociation reagent for 5 min at 37 1C.…”