Planning on doing some experiments between two cultures - C6 glioblastoma and astrocytes isolated from rat cerebral cortex (cultivated in same medium), to try and see what influence one has to the other (for example, if metabolites have an impact on C6 cells proliferation). Any suggestions on how to execute this idea? Mine is to use inserts, would be interesting to hear your ideas.
Thank you!
Inserts are one possible way to do this, you may want to consider encapsulating the cells in sodium alginate or methyl cellulose beads. By encapsulating the cell types in different beads you can look at soluble factors and cell responses to these factors quite nicely with the added benefit of A 3D matrix which has been shown to make the cells more active in terms of their release of soluble factors. If you are interested in this method I can give you more details.
Hope this helps
Andrew
Thank you, Andrew! Would be interesting to see the details on this one:)
so the scheme for this experiment is to passage the cells and spin them down aspirate off the old media. Then one can resuspend the cells in a relatively small volume of media (200 microliters). This can then be added to about 800 uL of 2% sodium alginate that has been filtered using a sterile filter. This mixture is then added drop wise to media that has a high concentration of Ca2+ for polymerization of the alginate. At this stage you can use a one mL syringe and variable amount of media that you add the drops into so as to form alginate beads with cells inside them that have different shapes so you can tell them apart for more specific analysis (1 mL of media for a flat disc shaped bead and 5mL for a relatively spherical bead. Let the beads polymerize for four minutes, then wash them 3x with the normal growing media. After washing, let the beads rest for 8 to 12 hours. Then using a sterile spoon or scoopula, you can transfer the different beads into a twelve well plate for co-culture.
Nice! Is it similar using methyl-cellulose? Because I've used it for spheroid formation, but not for the application of co-culture assays, just to measure the invasion of certain cells of one type.
I believe that it should work just as well with methyl-cellulose but I am not as sure as I have not worked with it much.
Andrew
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