Use of microcarriers in Mobius® CellReady bioreactors to support growth of adherent cells
© McGlothlen et al; licensee BioMed Central Ltd. 2013
Published: 4 December 2013
The Mobius® CellReady bioreactor product platform incorporates novel disposable technologies that provide optimal performance for suspension mammalian cell culture. Here we show the utility of EMD Millipore's 3L and 50L CellReady single use bioreactors for the cultivation of adherent mammalian cells on microcarriers. Cytodex3® and Solohill® collagen microcarriers were first tested in a mixing study to assess feasibility. We evaluated the normalized mixing speed required in the 3L and 50L to achieve a suspension of the microcarriers and enable growth of the cells.
Manufacturer specifications show Cytodex3® and Solohill® microcarriers to be similar in density and size. Working with this assumption, mixing studies where performed using the Cytodex3® microcarriers in 3L Mobius® CellReady and Solohill® Collagen coated in 50L single use bioreactor to determine the slowest agitation speed or the just suspended mixing power inputs (P/V)js, required to fully suspend the microcarriers so that the beads are equally distributed in the bioreactor.
Microcarrier distribution was assessed by sampling the bioreactor at varying depths. Then the dry weight of the microcarrier was used to determine the% relative sample weight to the target weight.
Cultured MDCK cells on Cytodex3® microcarriers grew to a peak cell density of ~1e6cells/mL using a power input of 0.6W/m3 with a 2L working volume after 3 days.
Cultured hMSCs on Solohill® microcarriers grew to a maximum total cell number of 6e6 cells using power input of 0.6-0.8W/m3 with a 2.4L working volume after 12 days.
Data from the mixing experiments demonstrate the just suspended mixing power input was determined to be ~0.6W/m3.
Cell growth experiments with hMSCs demonstrate comparable cell growth in the 3L and 50L Mobius® CellReady bioreactor with total number of hMSCs reaching 4e8 and 9e9 cells after 12 days at a agitation power input of 0.6-0.8W/m3
Initial cell growth experiments with adherent MDCK cells demonstrate an agitation power/volume input of 0.6W/m3 may provide the best performance for cell growth with peak cell densities ~1.0e6 cells/mL after 3 days
Comparable MDCK cell growth is observed:
Mobius® CellReady Bioreactor 3L
Mobius® CellReady Bioreactor 50L
Physical Characteristics of Microcarriers
Cytodex 3 ®
Solohill® Collagen Coated
Hydrated Size (µm)
MDCK/ Cytodex 3 ® Microcarriers Process Table
Cytodex 3 ®
DMEM w/ 4.5g/L Glucose, 2% FBS, 1% NEAA and 2mM L-Glutamine
DMEM low glucose, 10% FBS, 8ng/ml bFGF, 2mM Glutamine, 1X Pen/Strep
DO (% Saturation)
Day 1: 100% Growth Media
Day 6: 1000ml low glucose fresh medium
Day 3: Drain 50% of the working volume and reefed with equal volume of Growth Media
Day 9: 400ml high glucose fresh medium
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