I am researching on some gene expression dynamics in mouse muscle progenitor cells. There is one gene that has function in cell-cell communication. It is expressed in one cell and as a ligand to the neighbour cells. The neighbour cells also express this gene. When this gene is modified (not knock-out or knock-down), the dynamic of expression is changed. There have already been some similar reports in neural system and other tissue. But if they use isolated cell and single cell imaging, the dynamics in mutant are similar to the wild type. It means the cell-cell interaction may affect the dynamics. Now I want to check the situation in muscle system. I tried with mouse E12 embryo slice culture several times. But the progenitor cells grow and differentiate too fast. It is very hard to tracing. I also want to use cell mass to keep the interaction. I need a good ex vivo method to perform the experiment. Any expert has any idea? Thanks.
Hello Zhang.
If the local signal is juxtacrine one, it would be better to have an assay system that has both close cell-cell interactions (cell contacts) and isolated cells to demonstrate simultaneously activated or inactivated cells together in a culture dish. This system can be established only by replating appropriate cell numbers as a monolayer cell culture after dissociating the embryonic myoblasts. Delta-Notch signaling can be studied by this approach more closely.
Best wishes.
Hello
Yes , i agree with Sang Ho Lee , the monolayer cell culture is the good way in this case .
Kindly look at this article :
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3638420/
Hi Zhang.
It cannot be said due to the limited info. So I do not know the properties of your activated molecules.
Read it carefully "This system can be established only by replating appropriate cell numbers as a monolayer cell culture after dissociating the embryonic myoblasts."
So test your cell number by plating dissociated cells in varying cell concentrations. To establish such a screening system, you've got do such a little test run. You do not need the whole isolated myoblasts, take a proportion of cells for a test run to determine what cell concentration is appropriate within what intervals (in 16h or in 24h because they will proliferate greatly). Of course, this has to be GFP fusion to be traced by fluorescence microscope. If you put more info RG members will give solutions in no time.
Regards.
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