I am generating brain organoids with two different iPSCs cell lines. I have made many times brain organoids with one of these. However, when I start an experiment with other iPSCs cell line, some organoids (majority) have circular protrusion. What could be the reasons?
I could imagine that you have some choroid plexus cells in your organoids that produce some kind of cerebrospinal fluid. I have also experienced this from time to time but always very small bubbles < 10% of the organoid size and not so big ones. This might also depend on the protocol you use.
I agree with Nicole; it's the choroid plexus.
https://www.stemcell.com/products/stemdiff-choroid-plexus-organoid-differentiation-kit.html
The efficiency of differentiation of iPSCs may vary between cell lines. Sometimes this is also affected by the Matrigel. Matrigel may have different compositions between different batches and induce choroid plexus generation. Sometimes this is specific to the iPSC line. The differentiation potential of iPSCs may vary between lines.
Brain organoids are generally classified based on morphology for further study. Alternatively, you can compare those that spontaneously developed the choroid plexus with those without.
Experiments on iPSC-derived organoids require many repetitions. Unfortunately, there may be large differences between individual iPSC lines and between the individual organoids obtained from each of them. Moreover, large differences may also occur between differentiation experiments in the case of one iPSC lineage.
Good luck!
I agree with Nicole; it's the choroid plexus.
https://www.stemcell.com/products/stemdiff-choroid-plexus-organoid-differentiation-kit.html
The efficiency of differentiation of iPSCs may vary between cell lines. Sometimes this is also affected by the Matrigel. Matrigel may have different compositions between different batches and induce choroid plexus generation. Sometimes this is specific to the iPSC line. The differentiation potential of iPSCs may vary between lines.
Brain organoids are generally classified based on morphology for further study. Alternatively, you can compare those that spontaneously developed the choroid plexus with those without.
Experiments on iPSC-derived organoids require many repetitions. Unfortunately, there may be large differences between individual iPSC lines and between the individual organoids obtained from each of them. Moreover, large differences may also occur between differentiation experiments in the case of one iPSC lineage.
Good luck!
Thank you Burak for starting this "Question" and Nicole and Justyna for answering it! We're having the same problem here in our lab. This was very helpful!
We made very good experiences with Matrigel-independent protocols. Usually this organoids are more homogenous. If you want to avaoid choroid plexus formation in the future you might also try region specific organoid protocols. If you need suggestions I could help you there too.
Nice!
Thanks for your availability.
We'll certainlly come to you when we needed!
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