I wanted to perform LIVE /DEAD staining of Adipose derived Stem Cells (ASC) on the electrospinned Poly-L-lactide co-e-caprlatone(PLCL) nanofibers. 1 m2 PLCL nanofiber (collagen coated) was placed on each well of 24 well plates. ASC was prepare in aliquot of media and this was added to the PLCL nanofiber for the attachment of the cell for 3 hrs. This was followed by adding the remaining media in each plate. The problem that I encounter is that the PLCL floats on the media due to which the seeded cells proliferates to the bottom of the well plate rather than the nanofiber surface.
How can this problem be overcome?
Do I need to switch over to the special type of well plate that holds the nanofiber at the bottom thus avoiding it to float?
Or Is there any techniques by which I can perform the staining of ASC in the routine 24 well plate?
Dear Kshitiz,
To overcome floating nanofibrous material in 24 well plate you can use this:
http://www.sigmaaldrich.com/catalog/product/aldrich/z681903?lang=pl®ion=PL
I think, that you must use some kind of holding device for your material. You cannot just switch to other technique of staining, because your cells will never grow on the material if you will not hold it down. However, there might be another protocol for cell culture on nanofibrous material, other than with inserts, but I am not familiar with it.
When you use the insert you can use all protocols commonly reported in literature for staining the cell (for instance Live/Death).
Best regards,
Michał W.
Dear Michal,
Thank you for the suggestion. As per your advice, I will try the cell seeding in the PLCL, with the plate insert.
Also, you can prepare some inserts by yourself. You just have to use a material resistant to high temperature in order to sterilize the insert.
- Badges
- Science topic
- Similar topics
- Biological Science
- Biochemistry
- Biomolecules
- Peptides