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Useful Numbers for Cell Culture. Surface. Area. (cmA2). Seeding. Density. Cells at. Confluency. Versene. (mlof. 0.05%. EDTA). Trypsin. (mlof 0.0.5%.
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Materials § Culture vessels containing your adherent cells § Tissue-culture treated flasks, plates or dishes § Complete growth medium, pre-warmed to 37°C § Disposable, sterile 15-mL tubes § 37°C incubator with humidified atmosphere of 5% CO 2 § Balanced salt solution such as Dulbecco’s Phosphate Buffered Saline (DPBS), containing no calcium, magnesium, or phenol red § Dissociation reagent such as trypsin or TrypLE™ Express, without phenol red § Reagents and equipment to determine viable and total cell counts such as Countess® Automated Cell Counter, Trypan Blue and hemacytometer, or Coulter Counter® (Beckman Coulter) Procedures
Materials
Surface Area (cmA2) Seeding Density Cells at Confluency Versene (mlof 0.05% EDTA) Trypsin (mlof 0.0.5% tryspin 0. mM EDTA) Growth Medium (ml) Dishes 35mm 9 0.3 x 10 6 1.2 x 10 6 1 1 2 60 mm 21 0.8 x 10 6 3.2 x 10 6 3 2 3 100 mm 55 2.2 x 10^6 8.8 x 10^6 5 3 150 mm 152 5.0 x 10^6 20.0 x 10^6 10 8 Culture Plates 6 - well 9 0.3 x 10 6 1.2 x 10 6 2 2 3 - 5 12 - well 4 0.1 x 10 6 0.4 x 10 6 1 1 1 - 2 24 - well 2 0.05 x 10^6 0.2 x 10^6 0.5 0.5 0.5 - 1. Flasks T- 25 25 0.7 x 10^6 2.8 x 10^6 3 3 3 - 5 T- 75 75 2.1 x 10 6 8.4 x 10 6 5 4 8 - 15 T- 160 162 4.6 x 10 6 18.4 x 10 6 10 10 15 - 30 Reference: Life Technologies. Counting Cells in a Hemacytometer. Retrieved from http://www.lifetechnologies.com/us/en/home/references/gibco-cell-culture- basics/cell-culture-protocols/counting-cells-in-a-hemacytometer.html Life Technologies. Freezing Cells. Retrieved from http://www.lifetechnologies.com/us/en/home/references/gibco-cell-culture- basics/cell-culture-protocols/freezing-cells.html