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soft agar assay - cells in the top agar cells adhere to the plates unexpectedly. (Aug/31/2010 )

Hi, everybdy!
I've done soft agar assay for several times. However, I've got a trouble that cells in the top agar cells adhered to the plate unexpectedly a few days after I seeded the cells .

The following was My protocol.
1. Prepare 0.5% base agar layer and store the plate at 4C.let it sit at room temp for 30 min before using.
2. Prepare 0.6% top agarose solution
3. Harvest and count cells
4. Respend the appropriate number of cells in top agar . Final concentration of top agarose solution was 0.3%.
5. Incubate for 10-30 days and feed cells 2 times a week.

Did anybody have experiences?
What's the reason for the phenomenon that cells in the top agar cells adhered to the plate unexpectedly?

-zhu-

<_<
Nobody konw?

-zhu-

what sort of cells you are using?

-adrian kohsf-

Thanks for your reply.
I use murine barrow marrow mesenchymal stem cell.
Do you have any suggestions?

-zhu-

Hello Zhu, now I am experiencing similar problem i.e. cells in the top agar cells adhered to the plate. Did you find out the solution? Please let me know

-Manish Neupane-