Falcon Cell Culture Multi-Flask An easier, more productive way to culture cells

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1 Falcon Cell Culture Multi-Flask An easier, more productive way to culture cells Falcon Cell Culture Multi-Flasks enable you to grow more cells faster and easier, thereby making your cell culture workflow more productive. The tissue culture-treated Falcon Multi-Flasks are available in 3- or 5-layer formats that provide 525 cm 2 or 875 cm 2 cell growth surface area. They can be used with a wide range of liquid volumes (up to 50 ml/layer). Features Even distribution of media across all layers for homogeneous cell populations. Ability to mix in the Falcon Multi-Flask saves time and reduces risk of contamination. Flexible design lets you pour or aspirate/recover cells using a pipet. Consistent surface treatments for predictable scale-up. Manufactured in compliance to cgmp standards. Designed to Fit Your Protocol Simplify your scale-up. Falcon Multi-Flasks, offer the same footprint, the same reagent volumes, and the same cell seeding densities as 175 cm² flasks. Plus, you ll be using the same proven surface treatment as all other Falcon flasks. Improves Your Cell Culture Productivity Falcon Multi-Flasks deliver a thoughtful design that simplifies your workflow. You can eliminate multiple steps and reduce the risk of contamination. Pipet access allows you to aspirate media to exchange with fresh media and recover cells without pouring. The mixing port allows rapid mixing within the Falcon Multi-Flask to create your cell suspension, transfect your cells, or add other reagents directly to the flask. The port also provides for a uniform distribution of media and cells to facilitate homogeneous cell growth on all layers. More Consistent Cell Growth Falcon Multi-Flasks even distribution of media, proven vacuum-gas tissue culture surface treatment, and effective gas exchange all combine to provide an optimal cell culture environment. The result is higher cell yield and a more homogeneous cell population. Visit for more information. FALCON

2 PREDICTABLE SCALE-UP Cell Number x Cell Number x Corning T175 Flask (1-layer) Falcon Multi-Flask (3-layer) CELL CULTURE VESSEL Falcon Multi-Flask (5-layer) 0 BHK-21 LnCap Hep-G2 EcoPack CELL LINE Expected Yield Actual Yield T-175 Falcon Multi-Flask (3-layer) Falcon Multi-Flask (5-layer) Three and five times more BHK-21 cells were grown and recovered from 3- and 5-layer Falcon Multi-Flasks than T-175 flasks (left). Cell yields per cm 2 were equivalent in 3- and 5-layer Falcon Multi-Flasks and T-175 flasks for BHK-21, LnCap, Hep-G2, and EcoPack cells (right). CONSISTENT CELL PHYSIOLOGY CHO-M1 cells were grown in a T-175 and a 3-layer Falcon Multi-Flask for 72 hours. Cells were harvested and re-seeded on 96-well PDL plates for GPCR Calcium mobilization assay. Cells were challenged with M1 agonist, Carbachol (300 µm) and responses were recorded 1 minute following compound addition. Data is expressed as fold-increase in calcium flux compared to control cells challenged with assay buffer (no drug). No significant difference in agonist response was observed between cells harvested from either vessel type. Data shown is representative of three independent experiments. Calcium Mobilization Fold Increase Over Control (mean + SD) T-175 CELL CULTURE VESSEL Falcon 3-layer SIMPLIFIED WORKFLOW Pipet access eliminates the need for pouring. It also facilitates the use of a wide range of liquid volumes (up to 50 ml/layer), efficient recovery of valuable cells and reagents, and minimizes the risk of contamination. Mixing port eliminates mixing outside the vessel, saving you time and effort. This unique in-vessel mixing allows for rapid and uniform distribution of cells and reagents. Compatibility with similar volumes of reagent usage and cell seeding densities as T-175 flasks makes scaling up a familiar and simple process.

3 CONSISTENT CELL GROWTH This figure illustrates homogeneous cell growth between layers of Falcon Multi-Flasks. BHK-21 cells grown to >80% confluence in 3-layer Falcon Multi-Flasks and control T-175 flasks were fixed and stained with crystal violet. Control flasks and individual layers of the multi-flask vessels were cut and scanned. Top Middle Bottom Corning T-175 (control) IDEAL FOR PLURIPOTENT STEM CELLS Human Mesenchymal Stem Cells were cultured in MSCGM medium (Lonza) and seeded at 4,000 to 5,000 cells/cm 2 in a T-175 Falcon Flask and a 3-layer Falcon Multi-Flask. The cells were cultured until they reached 80% confluence (~6 days). Cells were harvested, counted, and cumulative population doublings of hmscs from each vessel type were calculated over five consecutive passages. Immunophenotypic properties of hmscs grown in the Falcon Multi-Flask as determined by flow cytometric analysis (FACS) confirmed that hmscs were negative for surface markers CD14, CD34, CD45, CD79a, and HLA-DR and positive for expression of surface markers CD73, CD90, and CD105. Respective isotype controls for each marker antibody are shown in grey. COMPATIBLE WITH YOUR CELL LINES Diverse cell lines and primary cultures (with and without serum) can be grown and efficiently recovered from Falcon Multi-Flasks. Expected yield was determined using average cell yield from control T-175 flasks multiplied by three and five times for the 3- and 5-layer Falcon Multi-Flasks, respectively.

4 Falcon Cell Culture Multi-Flask An easier, more productive way to culture cells PRODUCT SPECIFICATIONS WORKING VOLUME RANGE >5 ml per layer for dissociating <50 ml per layer for cell expansion MOLDED-IN GRADUATIONS 0 to 50 ml per layer in 10 ml increments GRADUATION ACCURACY 10% CAP VENT MEMBRANE 0.2 μm hydrophobic membrane CELL GROWTH SURFACE Tissue culture-treated, optically clear ORDER INFORMATION Falcon Cell Culture Multi-Flask Description Qty/Pack Qty/Case Cat. No. 3-LAYER TISSUE CULTURE-TREATED, 525 CM LAYER TISSUE CULTURE-TREATED, 875 CM To place an order in Europe, contact Customer Service at: tel: , fax: ; CSEurope@Corning.com For technical assistance, contact Technical Support at: cctech@corning.com ACCESSORIES Description Qty/Pack Qty/Case Cat. No. Falcon Pipets ASPIRATING PIPET 2 ML INDIVIDUALLY WRAPPED ML INDIVIDUALLY WRAPPED PAPER-PLASTIC ML INDIVIDUALLY WRAPPED PLASTIC-PLASTIC Falcon Tubes 50 ML POLYPROPYLENE CONICAL TUBE ML POLYPROPYLENE CONICAL TUBE ML POLYPROPYLENE CONICAL TUBE U.S. Orders Contact your authorized distributor to place your order. Corning acquired the Discovery Labware Business including the Falcon brand. For information, visit For a listing of trademarks, visit us at All other trademarks are property of their respective owners. Corning Incorporated, One Riverfront Plaza, Corning, NY Corning Incorporated Life Sciences Corning BV Fogostraat LJ Amsterdam The Netherlands Phone: Fax: cceurnl@corning.com , 2013 Corning Incorporated Printed in Europe 02/13 CLS-DL-CC-024 A4

5 Falcon Multi-Flask Frequently Asked Questions FALCON Why does media spill over to the other layers when moving the Falcon Multi-Flask in and out of the incubator? Transport the flask according to the user guide (see step 8) using the partition position and keeping fluid away from the mix port. Do not carry Falcon Multi-Flasks in a horizontal position as this may cause spillover of media between layers. Why does media distribution become uneven when I place the Falcon Multi-Flask in the incubator? Fluid may migrate from one layer to the other. To prevent this from occurring, one should: Ensure the incubator shelf is level. Position flasks across shelf without blocking the vented neck region. Ensure weight is distributed evenly throughout the shelf and the shelf can withstand the weight of flasks with media. Use the opposite corner of the mix port to pivot the flask and lay it horizontally. Ensure media does not flow back into the mix port or over the shelves into the neck region. I followed the user guide and set the Falcon Multi-Flask into the incubator. A few hours later, I see uneven media distribution by layer. What do I do? Repeat re-equilibration and partitioning steps (user guide steps 7 and 8). Ensure the work surface is flat while equilibrating and partitioning fluid in the Falcon Multi-Flask. Inspect the flask at least 2 hours after seeding. Re-equilibrate and partition again, if necessary. Minimize vibration on, or near, the surface where Falcon Multi-Flasks are housed. What volume of media should I use? Begin with the same media volume and cell seeding density on a per unit surface area basis as your existing device. The surface area of the flasks are 525 cm 2 for 3-layer and 875 cm 2 for 5-layer flasks. The maximum amount of media used should not exceed 50 ml per layer. What volume of dissociation reagent should I use? We recommend 5 ml per layer of Falcon Multi-Flask. Can I use the same volume of reagents as T-175 flasks? No, the amount of media per unit area should be the same not the actual total volume. If you currently use a T-175 flask, simply multiply the volume by 3 or 5 times depending on the Falcon Multi-Flask format used.

6 Why do I observe excessive frothing or bubbling of media and can this affect my cell culture? Over mixing can result in excessive frothing or bubbling of media, which can lead to fluid migration from one layer to another. To avoid this, follow user guide step 5. While the flask is in the mixing position, allow the media to drain only from the layer on top; immediately tilt the flask in the opposite direction; and repeat. Ensure the flask is brought back to the mix position (user guide step 4) before equilibration of media. Improper mixing or pouring of media into the flask can also create foaming or bubbles. The correct way to add media is to follow step 1 in the user guide and pour or pipet gently along the slope of the flask lid. How can the flasks be stacked in an incubator? Ensure the flasks lock into position using the stacking ribs. Position the stacks of flasks evenly in the incubator to prevent the shelves from bowing. Allow adequate room for air circulation through the vented cap of the flasks. When stacking the Falcon Multi-Flask on incubator shelves that are above shoulder height, ensure the media distribution by layer remains equal. The use a step-stool can help facilitate flask placement. What is the best way to add cells to the vessel? Add the desired volume of growth media to the flask and keep it in an upright position. Add cell suspension stock (at least 10% final volume) directly into the growth medium using a 10 ml pipet. Is pre-wetting of the flask or pre-warming of the media in the flask required? Neither is required, nor are they recommended unless you are applying a coating to the flask growth surface. The 5-layer Falcon Multi-Flask will not fit under my microscope. What do you recommend? With some inverted microscopes, the condenser can be removed or moved to accommodate the 5-layer flasks. If this is not possible, a control flask (T-175) can be run side-by-side to provide an indication of cell morphology and confluence. Why is cell growth patchy when I view the Falcon Multi-Flask under the microscope? Uneven cell/media distribution most likely occurs from inadequate mixing. Refer to steps 4 and 5 of the user guide for the proper procedure. Don't use a clumpy cell suspension stock. Ensure cells are adequately triturated to generate a single cell suspension. Why am I getting media in the neck of the flask? Media should not enter the neck of the flask if the Falcon Multi-Flask is used properly. Check to ensure the flask is not upside down. The logo should face up when the flask is laid horizontally for incubation. Over-rotation when mixing cells in the vessel can result in media getting into the neck area of the flask. To avoid this, please follow steps 4 to 6 in the user guide. Why are my cells not attaching? If your cells normally attach to a single layer flask, then they will attach to the surface of the Falcon Multi-Flask. Ensure the flask is laid in the correct position with the logo facing up. In this position, cells will adhere to the tissue culture surface on the base of each layer.

7 Can I pour my cell suspension into the flask? Yes, however we recommend that to avoid foaming of media, use a 50 or 100 ml pipet (Cat. Nos and ). Insert the pipet past the logo and stream liquid along the top slope of the flask into the first layer. Can I pour my cells and reagents out of the flask? Yes, you can pour or use a pipet to recover the cells during harvesting. If cells clump, mechanically dissociate the cells by titrating the suspension. How can I maximize the recovery of cells and reagents? To minimize residual volumes, pour as per step 13 in the user guide. Allow liquid to pool on the mix port side prior to pouring with the logo down. For maximal recovery of cells and reagents, follow steps 11 and 12 in the user guide. First, aspirate by positioning the pipet tip towards the mix port region. Then, pool liquid into the top layer and tilt the flask to partition liquid away from mix port. For high density cell cultures, additional rinses are recommended. Visually inspect the base layer to determine how many rinses are required. Why am I not getting the expected cell yield? You may not be recovering all the cells. Additional rinses may be required, as noted above. You may have uneven distribution of cells and media. See the user guide for proper mixing, equalization and partitioning procedures. You should avoid lengthy delays between cell seeding and incubation so that cells do not settle to the bottom of the vessel.

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