Last Updated 11/1/07 MDR
MEDIUM
The recommended ratio of medium volume to surface area for monolayer cell cultures is 0.2-0.3 ml/cm2 *. The upper limit is set by gaseous diffusion through the liquid; the optimum ratio depends on the oxygen requirement of the cells. Cells with a high oxygen requirement grow better in shallow medium (~2mm), those with a low requirement grow better in deep medium (~5mm). Exceeding a medium depth of 5mm can hinder gas exchange and impair growth.
*96WP requires a ratio of 0.3-0.4 ml/cm2 to overcome surface tension's effect on cell distribution and growth in the wells
*Volumes used in tissue culture flasks are limited by the locations of the neck
TRYPSIN
PD: petri dish
WP: well plate
T: tissue culture flask
| Dish | Area (cm2) | Medium Volume (ml) | Trypsin-EDTA Volume (ml) |
|---|---|---|---|
| PD15 | 176.7 | 35-50 | 5-7 |
| PD10 | 78.5 | 15-24 | 2-3 |
| 6WP | 9.6 | 1.9-2.9 | 0.40 |
| 12WP | 3.8 | 0.760-1.14 | 0.15 |
| 24WP | 1.8 | 0.360-0.540 | 0.075 |
| 48WP | 0.75 | ||
| 96WP | 0.20 | 0.060-0.080 | 0.01 |
| T25 | 25 | 5-6 | 1 |
| T75 | 75 | 15-20 | 2-3 |
| T175 | 175 | 35-50 | 5-7 |