I'm currently working with mouse uterine organoids and I've seen that many authors perform organoid passaging using mechanical or enzymatic methods to dissolve the Matrigel while breaking the organoids to a single-cell suspension.
I'm wondering, what's the point of breaking organoids to single-cell suspension while passaging?
I understand the passage as a method to allow the organoids to have more space to grow; to replace the expired Matrigel; and to get rid of uninvited cells that are growing in monolayer and are consuming the organoid media. However I don't understand why I shoud break the organoids if my objective is to allow their growth in number and size.
Thank you.
Dear Dr. Federica Marinaro
I don't understand why I should break the organoids if my objective is to allow their growth in number and size?
Organoids must be passaged because if they are too large (typically bigger than 500μm), they become hypoxic and necrotic due to lack of oxygen and nutrients in the core of organoids. Thus, it is essential that you control the size of organoids by physical and enzymatic dissociation for reproducible generation of data from organoids.
What's the point of breaking organoids to single-cell suspension while passaging?
Another important point to be noted is that during passaging, it is critical that the organoids are not broken up too small or else the organoid formation efficiency will be drastically reduced. Your goal should be to break each organoid into multiple fragments, not to reduce organoids to the single cell level. I would not recommend breaking organoids to single-cell suspension.
Regards,
Malcolm Nobre
- Badges
- Science topic
- Similar topics
- Psychology