Hi all,
I am a physical chemistry PhD student currently working with 3D cell culture on polycaprolactone UNTREATED scaffoldings, for NMR and MRI purposes only.
I am not particularly interested in cells physiology, althouh I understand this is important to optimise cell adhesion on the scaffolds I plan to do imaging on.
I am working with mesenchymal stem cells, which differentiate into osteoblasts after the seeding.
My scaffolds are disc shaped, 1.5 mm thick, kept in 96-well plate. I seed each scaffold with a 20 uL drop of cell suspension, topping up the wells with media after 3-5 h of incubation.
I do know I can make the scaffolds more appealing to the cells by coating them.
However I wondered:
- can I make the bottom of the plate LESS appealing for the cells, in order to convince them to attach to the scaffold from the get-go?
- would that damage the cells, as they sink to the bottom for gravity and they find an unconfortable environment to attach to? and
- could that be avoided by keeping the suspension gently agitated so that cells have more chances to come in contact with the scaffold?
I tried UNTREATED wells too, but cells seem to like those anyway, no difference with the treated wells.
Many thanks
Giulia
There are silanes, some of them available in aqueous solution that can be used to coat the dish/well and reduce cell adhesion. They might interfere with adhesion of your scaffold to the dish, however. Article Micropatterning of Au NPs on PEG Hydrogels Using Different S...
https://www.thermofisher.com/order/catalog/product/TS-42799
https://www.scientificlabs.co.uk/handlers/libraryFiles.ashx?filename=Manuals_L_LZ50640_A.pdf
there are low cell attachment plates available from many companies used for Organoi/spheroid formation, but they tend to be round-bottomed, not great for some microscopy applications.
Giulia Melchiorre You can try agarose coating of the wells before loading the scaffold. It worked for me. 1.5% hot solution casting of 40ul will be enough for 96 well plate.
You may want to try coating the well with agarose before adding the scaffold. And add an adhesive ligand like collagen to the scaffold to increase adhesion on the scaffold. I use this and it works.
Giulia Melchiorre Siliconising agents as the ones suggested by Ellen A G are most suited for glass surfaces. For plastic surfaces I prefer this product: Anti-Adherence Rinsing Solution for Cultureware | STEMCELL Technologies. Extremely easy and quick to apply!
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