Application Note� ��������������
SuperSpinner S / SuperSpinner C
Upscale cultivation and proliferation of bovine aortic endotheleial cells.
Combination of the SuperSpinner with dextrane based microcarriers.
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.
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
- 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
- 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
- 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
Schwarzenberger Weg 73–79 Fax +49 (0) 5661.71.370234212 Melsungen
Sartorius BBI Systems
No. 12-01 Date: January 10, 2012 Bureau of Emergency Medical Services Re: Blood Glucometry and Nebulized Albuterol POLICY STATEMENT for EMS Agencies Supersedes/Updates: 09-13 Page 1 of 2 BACKGROUND The New York State Emergency Medical Advisory Committee (SEMAC) has approved the use of glucometers and nebulized albuterol by Emergency Medical Technicians (E
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