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Application Note�������������
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SuperSpinner S / SuperSpinner C
Upscale cultivation and proliferation of bovine aortic endotheleial cells.
Combination of the SuperSpinner with dextrane based microcarriers.
Original article:
Müthing, J., Duvar, S., Nerger, S., Büntemeyer, H. and Lehmann, J. „Microcarrier Cultivation of Bovine Aortic Endo-
thelial Cells in Spinner Vessels and a Membrane-stirred Bioreactor.“ Cytotechnology 18 (1996): 193-206.
Cell types
A basic adaption protocol is provided for cultivation of attachment dependent primary endothelial cells grown on Goal of cultivation
microcarriers in a membrane aerated spinner bioreactor. Reliable proliferation of sensitive adhesion dependent cells Subsequent adaption of medium volume, stirrer speed in larger scale systems. Comparison of different surface and medium supplementation/exchange allowed a fast qualities of dextrane based microcarriers.
and effective proliferation (factor 4 in eight days) and maintenance with high viability (>90%) for 32 days.
Basic culture conditions
- 2l BIOSTAT® BF bioreactor with membrane aeration
Further hints
- DMEM (4mM glutamine, 25 mM glucose), supplemented The used microcarriers are no longer commercially available. with 15% newborn calf serum, 44 mM NaHCO Different products can be used instead. Macroporous sodium pyruvate, 65 mg l-1 benzylpenicillin, 100 mg l-1 microcarriers may result in limited oxygen supply.
(iA. Preismann, R. Wiesmann, R. Buchholz, R.G. Werner, W. Noe. - Microcarrier: Dormacall®, dextrane matrix beads „Investigations on Oxygen Limitations of Adherent Cells Growing on covalently coated with positively charged dimeric N, Macroporous Microcarriers.“ Cytotechnology 24 (1997): 121-34.
N-Diethylaminoethyl-groups- Cultivation Temperature 37°C- Aeration: 50% O2, 10% CO2 - Rotation speed of membrane stirrer bar: adapted to cultivation phase (s. below) Inoculation and attachment phase, proliferation and
productivity
- Initial cell density: 4x105 ml-1
- Medium volume: 500 ml
- Microcarrier: 6g (~37.2 x 106 cells) total
- Rotation speed of membrane stirrer: 30 rpm for 2 min in
intervalls 20 min within the fi rst 12 hours, afterwards
30 rpm continuously until hour 48
Proliferation and steady state phase (day 2-10 and 10-32
respectively)
- Medium volume: 2000 ml
- Rotation speed of membrane stirrer: 50 rpm
- Glutamine supplementation on day 13
- Medium exchange on day 20
Sartorius BBI Systems GmbH Phone +49 (0) 5661.71.3400 Engineering by
Schwarzenberger Weg 73–79 Fax +49 (0) 5661.71.370234212 Melsungen Sartorius BBI Systems

Source: http://www.sartorius.or.kr/nh/fileadmin/sartorius_pdf/fermentors-bioreactors/application_note_4_221104_microcarrier.pdf