After 21 days of differentiation the pellet will be harvested for histology & immunohistochemistry, What is the best way to harvest the pellet without affecting the cell morphology?
Please someone should help
Thank you Antonia. Please I want to hear other peoples experience
You want the sedimentation force in earth gravity units (g), not rpm, since the rpm for a given g force depends on the radius from center of rotor to the tube bottom. For most mammalian cell cultures we use about 100 g to 200 g for 5 to 10 min at room temperature. This minimizes cell trauma.
Hello Suleiman,
I have done chondro differentiation and according to my experience centrifugation at 1000 rpm for 10 minutes (in 15ml falcon) is optimized method to maintain cell morphology as well as preventing cell death and facilitating optimum cell aggregation for chondro differentiation. We used a kit and 1k rpm fr 10 min worked quiet fine. You can easily change media in this pellet without going for further centrifugations. Maintain tubes in semi-tight state to allow gas exchange and change media every third day (in 15ml falcons only) like routine culture ..
Hope it helps !
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