i will test some drugs on the ability of forming tumorsphere with my cancer cell line. I know i will add the drug the next day of seeding to the media, but how to remove it after 3 days without disturbing the sphere formation ?
Tricky problem. What I've attempted in semi-similar circumstances was to carefully pipette out the majority of the media and then replace it, all the while observing the tumor sphere (if visible by eye) to ensure no disruption. Doing this a few times will significantly dilute the reagent or drug in question. However, if the tumor sphere fails to form strong intercellular connections, you can easily disrupt the sphere and even end up pipetting out single cells or small clumps of cells. An alternative might be to simply add media until the well is full, and then remove it, multiple times. This way you will still be diluting out the drug yet it might have less of a disruptive influence on your tumor spheres. Another alternative -you could use a wide-tipped pipette and actually remove your tumor spheres from the plate and re-plate them in drug-free media. Depending on the strength of the intercellular contacts (again), this may or may not work. Good luck.
Osama,
for this purpose we have used Corning ultra-low attachment plates to grow spheroids and Matrix Platemate system with 30ul tips. Spheroids remained in the well after removing 30 ul of 35 ul of media.
- Badges
- Science method
- Similar topics
- Biological Science
- Histology
- Staining