Corning

Q: What is the recommended culture medium working volume in a Corning 5L Erlenmeyer flask?A: We recommend a working volume range of 2.5 to 3.0L of culture medium with Corning 5LErlenmeyer flasks. Specifically for the baffled option, we recommend a working volume no less than 2.5L, the minimal volume for which the baffles were designed and optimized. Due to the height of the baffles, working volumes less than 2.5L could experience forces that take away the benefits of the baffles. Q: What is the recommended agitation rate for Corning 5L Erlenmeyer flasks?A: The optimal agitation rate will depend on the diameter of the orbit of the individual shaking platform, the cell type being cultured, the medium used and whether a baffled or plain bottom Erlenmeyer flask is used. With 25 mm orbit shakers, we recommend a rate of 90 rpm for CHO and Sf9 cells when cultured in Corning 5L Erlenmeyer flasks.It is important to empirically determine the agitation rate depending on conditions. This could be done by observing the amount of culture medium foaming with different agitation rates (90 to 120 rpm). The agitation rate with no or minimum foaming should be used as a good start-ing point. Typically we find the ideal rpm will need to be decreased as the size of the Erlenmeyer flask increases.Q: What is the recommended seeding density for 5L Erlenmeyer flasks?A: We recommend using the seeding density optimized for your given cell type or clone. Seeding den-sities of 0.3 to 0.5 x 106 cells/mL are typical for CHO cell culture and 0.5 x 106 to 1 x 106 cells/mL for Sf9 cell culture. Q: What is the recommended culture medium-feeding protocol in Corning 5L Erlenmeyer flasks?A: We recommend using the culture medium and feeding protocol (fed batch) optimized for your given cell type as a starting point. Some fine tuning of the feeding protocol may be needed for the maximum performance in 5L Erlenmeyer flasks. Q: What is the recommended scale up protocol for CHO cells in Corning 5L Erlenmeyer flasks?A: If starting from cryopreserved CHO cells, we recommend thawing cells into a Corning 125 mL Erlenmeyer flask using your standard protocol. Culture cells for several passages until cells are fully recovered and reach >2 x 106 cells/mL density by day 3 or 4. Always use cells from the mid logarithmic growth phase for the scale up. From 125 mL flask (40 mL working volume), scale up cells to 1L Corning Erlenmeyer flask (300 mL working volume), then to 5L Erlemeyer flask (2.5 to 3.0L working volume). It could take about 6 to 8 days to scale up from 125 mL to 5L Erlenmeyer flask depending on the cell growth rate. Q: Why might I want to use the baffled Corning 5L Erlnemeyer flask over a standard 5L Erlenmeyer flask?A: Usually we find that the baffles provide better aeration for cells, especially at a lower rpm. You may find that by using a baffled Erlenmeyer flask you can decrease the rpm from what you typically use.Q: Why is there an angled feature around the neck of the vessel?A: The angled feature located at the neck of the vessel will reduce foam when filling and pouring out of the vessel.Corning ® 5L Erlenmeyer FlaskFrequently Asked QuestionsFor a listing of trademarks, visit us at /lifesciences/trademarks. All other trademarks are the property of their respective owners.© 2014 C o r n i n g I n c o r p o r a t e d . A l l r i g h t s r e s e r v e d . P r i n t e d i n U .S .A . 4/14 C L S -B P -027Corning Incorporated Life Sciences836 North St.Building 300, Suite 3401 Tewksbury, MA 01876t 800.492.1110 t 978.442.2200 f 978.442.2476/lifesciencesWorldwide Support OfficesA S I A /P A C I F I CAustralia/New Zealand t 0402-794-347Chinat 86 21 2215 2888f 86 21 6215 2988Indiat 91 124 4604000f 91 124 4604099Japant 81 3-3586 1996 f 81 3-3586 1291Koreat 82 2-796-9500 f 82 2-796-9300Singaporet 65 6733-6511 f 65 6861-2913Taiwant 886 2-2716-0338 f 886 2-2516-7500E U R O P E Francet 0800 916 882f 0800 918 636Germanyt 0800 101 1153f 0800 101 2427The Netherlands t 31 20 655 79 28f 31 20 659 76 73United Kingdom t 0800 376 8660f 0800 279 1117All Other European Countriest 31 (0) 20 659 60 51 f 31 (0) 20 659 76 73L A T I N A M E R I C A Brasilt (55-11) 3089-7419 f (55-11) 3167-0700Mexicot (52-81) 8158-8400f (52-81) 8313-8589Warranty/Disclaimer: Unless otherwise specified, all products are for research use only. Not intended for use in diagnostic or therapeutic procedures. Not for use in humans. Corning Life Sciences makes no claims regarding the performance of these products for clinical or diagnostic applications.。