//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|
|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|