3D Culture of Mesenchymal Stem Cells

Example 1

A 3-D insert built in a 100 mm diameter non-adherent culture dish (Corning, Corning, NY) was used to culture human bone marrow-derived mesenchymal stem cells. Dimensions of the micro-channels were 3 mm width by 40 mm length by 10 mm height. 50,000 cells were seeded in each microchannel. Final volume of culture medium was adjusted to 400 uL. The culture dish was placed on a programmed rocking platform and cultured for 3 days with the setting at 5 rpm and 20° tilt. FIGS. 3A-3C shows cell aggregates at 24 hours, 48 hours, and 72 hours of culture. Aggregates were collected and replated on standard tissue culture plates for further culture (see FIGS. 3D and 3E).