I am growing ovarian cancer organoids and they grow very well. Sometimes the matrigel is dislodged from the bottom of the plate and I need to replate them. I follow a protocol where I wash the cells with cold D-PBS and then apply Corning's recovery solution to the organoids, place them on ice with mechanical shaking for 1 hr. Then I add trypsin for up to 5 min then I top up with RPMI, spin down, wash with DMEM, spin again and then use fresh matrigel on ice to re-plate.
My problem is that the cells do not seem to grow well after re-plating and the matrigel gets dislodged again from the bottom of the plates, any suggestions on how to solve this issue?
Hi,
In my opinion, you should change the method of ovarian cancer organoid embedding in the extracellular matrix.
You should embedding individual aggregates in the undiluted cold Matrigel. After polymerization ovarian cancer organoids should be cultured in an incubator on a shaker as a floating 3D culture. In this situation, replate them will be not necessary.
I think that Corning® Cell Recovery Solution can be used for the collection samples but not for passaging. To my knowledge, ovarian cancer organoids couldn't be passaging.
Best,
Justyna
Justyna Augustyniak thank you! So you mean that normally after the matrigel polymerises it should not adhere to the bottom of the plate but should float in media? Do you have any publication on ovarian organoid passaging?
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