Hello everyone,
I have the following question regarding the number of organoids in culture.
I performed a seeding assay with mouse tumor colorectal organoids. I seeded 2000 cells (dissociated from the organoids) and take pictures after 3 and 5 days. And then count the number of the experimental and control.
I observed a difference but the question is, Why is this possible? If I seed the same amount of cells and from this individual cells the organoids will grow, why the number of formed organoids can be different?
Some cells from a forming organoid scape and grow independently to form another organoid? Or some cells from the 2000 seeded cells do not grow and die?
All your comments are welcome
Thank you in advance.
There are a few potential reasons why the number of organoids formed could be different between experimental and control groups, even when starting with the same number of seeded cells:
- Variable viability of cells - Some seeded cells may die off before forming organoids, reducing the final organoid count. If there are differences in cell viability between experimental and control groups, this could lead to different organoid numbers.
- Clumping of cells - Seeded single cells may stick together and form clumps, which then grow into a single organoid. More clumping in one group could reduce organoid counts.
- Fragmentation of organoids - As organoids grow, some may fragment into smaller organoids. More fragmentation in one group could increase organoid counts.
- Differences in organoid coalescence - Growing organoids may fuse together. Less coalescence in one group could increase organoid counts.
- Variable organoid growth rates - If organoids grow at different rates between groups, faster growing organoids may appear sooner and be counted, impacting numbers.
- Seeding technique inconsistencies - Small differences in seeding density or distribution could lead to differences in organoid development.
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