Surface Areas and Guide for Recommended Medium …

1 This Guide gives the Recommended Medium volumes, approximate growth Surface Areas , and average cell yields for Corning disposable cell culture vessels. Approximate growth Surface Areas are based on calculations made from engineering drawings. These calculations do not take into consideration minor variations that can occur in products during molding or the ability of many cell lines to grow up the sides of the vessels which can considerably increase the available Surface area. For critical work, we suggest that you fix and stain cultures, and then carefully calculate the actual growth Surface general, at least 1 x 105 cells/cm2 can be produced when growing cells as attached monolayers in culture.

2 The average cell yields used here are based on this number. Actual cell yields can easily be several times higher or lower than this depending on the cell line and culture optimal cell to Medium ratios is important for obtaining good cell growth. As a starting point, we recommend to mL Medium for each square centimeter of culture vessel growth Surface area; most of the Recommended Medium volume levels used in the tables below are based on this ratio. Medium volume recommendations for microplates and Transwell inserts are higher due to meniscus effects associated with very small spaces and a higher rate of evaporation. Using more Medium may reduce the need for feeding the cultures, but, due to the increased Medium depth and the static nature of the environment, it will also slow the diffusion of oxygen to the Areas and Guide for Recommended Medium Volumes for Corning Cell Culture Vessels 2 Corning Microplates Single Well Only Well Approx.

3 Total Well Working Diameter Growth Average Volume Volume Microplate (Bottom) (mm) Area (cm2) Cell Yield ( L) ( L)Corning 96-well MicroplatesFlat Bottom x 104 360 100 - 200 Round Bottom N/A** N/A** 330 100 - 200V-Bottom x 104 320 100 - 200 Half Area x 104 190 50 - 100 Corning 384-well MicroplatesStandard x * x 103 112 25 - 50 Low Volume x 103 50 5 - 40 Corning 1536-well MicroplatesLow Volume x 103 1 - Flat Bottom * x 103 5 - 10 Solid Flat Bottom * x 103 5 - 10*Square wells.**Because these wells are round, the Surface area available for cell attachment is dependent on the Medium volume Multiple Well Plates Single Well Only Well Approx.

4 Total Well Working Diameter Growth Average Volume Volume Plate (Bottom) (mm) Area (cm2) Cell Yield (mL) (mL)6-well x 105 - x 105 - x 105 - x 104 - Permeable SupportsTranswell Transwell Insert Approx. Growth Average Insert Format Diameter (mm) Area (cm2) Cell Yield Well Insert6-well 24 mm cm2 x 105 12 mm cm2 x 105 mm cm2 x 104 mm cm2 x 104 mm dish 75 mm 44 cm2 x 106 Dishes Approx. Growth Average Recommended Dish Area (cm2) Cell Yield Volume (mL)35 mm* 9 x 105 - mm* 21 x 106 - mm* 55 x 106 11 - mm* 152 x 107 - mm 500 x 107 100 - 150*Not actual bottom diameters.

5 Dish is Volume (mL)3 Corning Flasks** Approx. Recommended Growth Average Medium Volume Approx. Total Flask Area (cm2) Cell Yield (mL) Flask Volume (mL)25 cm2 25 x 106 5 - 70 rectangular75 cm2 75 x 106 15 - 265 U-shaped150 cm2 150 x 107 30 - 45 377 U-shaped175 cm2 175 x 107 35 - 513 U-shaped225 cm2 225 x 107 45 - 1,006 traditionalCorning HYPERF lask 1,720 x 108 560 - 565 560 - 565 **Corning flasks (larger than 100 cm2) are considered US Class I medical Stacked Chambers** Approximate Average Recommended Chamber Size Growth Area (cm2) Cell Yield Medium Volume (mL)

6 Corning CellSTACK Chambers1-stack 636 x 107 127 - 1912-stack 1,272 x 108 254 - 3825-stack 3,180 x 108 636 - 95410-stack 6,360 x 108 1,272 - 1,90840-stack 25,440 x 109 5,088 - 7,632 Corning HYPERS tack Chambers12-stack 6,000 x 108 1,30036-stack 18,000 x 109 3,900**Corning stacked chambers are considered US Class I medical Roller Bottles** Approximate Average Recommended Roller Bottle Growth Area (cm2) Cell Yield Medium Volume (mL)490 cm2 490 x 107 100 - 150850 cm2 850 x 107 170 - 2551,700 cm2 ESRB 1,700 x 108 340 - 5101,750 cm2 1,750 x 108 350 - 525**Corning roller bottles are considered US Class I medical CellCube Systems**CellCube Approximate Average Recommended Module Growth Area (cm2) Cell Yield Medium Volume (mL)10-stack 8,500 x 108 N/A*25-stack 21,250 x 109 N/A*50-stack 42,500 x 109 N/A*100-stack 85,000 x 109 N/A**Not applicable; these systems are perfused with Medium from a reservoir.

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