Microcarrier-based shaken bioreactor: a convenient platform for achieving high cell density of anchorage dependent cells

Microcarrier-based bioreactors provide scalable and controllable systems to produce high doses of the cells required for clinical trials. Orbitally shaken bioreactors have been regarded as favorable reactors for mammalian cell culture as they allow sufficient oxygen and nutrient transfer at relatively low power consumption. Our main goal is to develop a microcarrier-based shaken bioreactor for culturing anchorage dependent cells.MethodsFirst, the internal surface of the bioreactor was silliconized using the solution of 2% (v/v) dichlorodimethylsilane in chloroform to avoid cell attachment to the bioreactor walls. Before cell inoculation to the microcarriers (Cytodex 3), they prepared according to the manufacturer’s instructions which contains washing with PBS, streilization and incubation in the medium. The bioreactor was cylindrical (total volume 250 ml) and was filled with 25 ml of the medium containing 5 g/L Cytodex 3. MC3T3-E1 cells inoculated onto the microcarriers at a density of 100000 (cells/ml) with intermittent shaking of 27 min off, 3 min on at 90 rpm (just suspended agitation rate) for the first 3 hours. After 3 hours the bioreactor was continuously shaken at 90 rpm. The cells were expanded in the bioreactor for 7 days with 50 % medium exchange every day. Cell proliferation was assessed on the day 1, 3, 5 &7 using the MTT colorimetric assay.ResultsOur bioreactor allowed microcarrier suspension as well as sufficient mass transfer without adverse effects on the cell viability and the microbeads integrity. Cells expansion on the microcarriers continued till day 7.ConclusionOur results demonstrated that orbitally shaken bioreactors provide an appropriate growth environment for long term of microcarrier-based cell culture

کلیدواژه ها:

Shaken bioreactor ، Microcarrier ، Cell culture