There are informations about centrifugation of cells before exchange a medium but I have a doubt if this procedure doesn/t damage spheroids?
under 600rpm for less than 5 minutes
This conditions are enough to centrifuge the typical spheroids.
Transfer your spheroids with media to a tube (15 or 50 ml). Let tube sit at room temperature for 30 min without any disturbances. All spheroids will go to the bottom of the tube. Remove most media by pipet or vacuum and leave 3-5 ml media in tube. Pipet up and down gently the 3-5 ml media containing spheroids and transfer it back to your culture dish with fresh media.
To aspirate the medium, I use ELx405 washer from BioTek. It works well for 96- and 384-well plates. Spheroids just need to be at the bottom of the wells, no centrifugation required. Then I just add fresh media with a multipipette.
Thank you for your advice. I have T98G glioblastoma multiforme cells.
Hello Anna,
if it is a short term experiment (7 days) then you can just add your culture with fresh media and no need to change the media as in some cases, cell number is a very crucial part and you can't afford to lose your cells or introduce unnecessary errors by centrifuging the spheroids in falcon tubes.
If it is a normal practice when you are culturing spheroids in T75 or T25 flasks and spheroid number is not of much importance you can always opt for tube method. Collecting the spheroid suspension in falcon (cut the tip end so as to get all spheroids in suspension of falcon), centrifuging it and then resuspending in fresh media and reseeding in culture flask but same practice can't be implemented during plate experiments like 96 well or 24 well. Because of my personal experience, I suggest you to centrifuge the whole plate using a plate rotor then, spheroids will settle down and then you can decant out the media gently and add fresh media. This will help to prevent any spheroid loss.
Hope it helps !
Hello Anna, we did devellop porous matrix based on hyaluronic acid that help to study spheroid where you'll be able to change medium as you want during more than 1 month culture. Let me know if you want to know more about it.
I work with cellstar plates for my spheroids (HCT8 cell line) and I have problems with refreshing media without sucking up the spheres.. Could anybody help me with this?
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