Application Note������������� Application Note� �������������� ����������� 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
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British Journal of Nutrition (2008), page 1 of 6Both inorganic and organic selenium supplements can decrease brainmonoamine oxidase B enzyme activity in adult ratsYa-Li Tang1, Shih-Wei Wang2 and Shyh-Mirn Lin1*1Department of Food and Nutrition, Chung Hwa University of Medical Technology, no. 89, Wenhwa 1st Street, Tainan County 717,Taiwan2Graduate Institute of Biological Science and Technology
Application Note�������������