Hi everyone,
When culturing a hydrogel in a well-plate, how do you decide how much medium to put over the gel? I have noticed that for 2D culture, the recommendations for medium volume give between a 2 to 3 mm height of medium over the cells (this is based on recommended working volume ranges given by Corning Life Sciences for their brand of cell culture plastics which I frequently use). Do you take in account how thick of a gel you are working with? How frequently will you change the medium?
Hello Sebastian,
Putting the media volume in well depends mainly on thickness of your gel and its volume. If you are using a thick gel then it should be completely covered into media and that media should be optimum so that it doesn't evaporate for two days. So you have to adjust that manually from your own experience. I must tell you that putting extra media will not cause much harm (but less media will definitely do) as cells will be properly embedded/adhered in gel. For a 24-well plate format and a volume of 200ul hydrogel, which just covered the bottom of well, I used to put 500-600ul media (which may vary upto 800ul if your gel is thick). I used to change media at every 3rd day and cells grew pretty well.
Hope it helps!
Sebastian, as Ajay said, make sure the hydrogel construct is submerged. The humidity in the incubator will help stabilize the depth of the pool. Excessive fluid will hinder pH regulation of the medium by the CO2 in the incubator. I have adhered to the Corning recommendation of 2-3mm over the top of the construct, changing half the medium every other day.
3D cell laden hydrogels must be completely immersed in the culture medium otherwise there will be no proper distribution and proliferation of cells in the upperside. Usually for my studies I used to change medium once in 2 days to estimate ECM secretions etc.
- Badges
- Science topic
- Similar topics
- Biological Science
- Microbiology