Previously, I used to seed primary dermal fibroblasts (which initially had slow growth rate) in collagen hydrogel and the cells used to show complete attachment and the normal shape of fibroblasts after culturing the gel in complete media. Now I have new fibroblasts which grow fast in flasks and once seeded in the hydrogel, they show some attachment within few hours but then start to take the circular shape and make colonies which make any further tests difficult.
I have tried different collagen concentrations (1, 1.5 and 2 mg/ml) as well as different cell densities (5000, 10000, 20000, up to 100000 cells per 50 and 100 microlitre hydrogels, but the behaviour of the cells were the same.
Please, could you explain how I can overcome that problem?
Dear Amir,
I have worked with these hydrogels but used stem cells. If you are putting cells inside the gel, they will die very fast and don't really proliferate inside the gel. I compared both methods.
The better way is to coat gel first on plate wells and then seed cells on it's top. They will still grow in 3D conditions. Cells really grow well in those conditions.
Hope it helps!
Dear Amir
I agree with the previous answer (Ajay Kumar), it is convenient to first treat the plate and then load the fibroblasts, I recommend a low density and a consentration of the hydrogel at a lower concentration
Thank you Ulises and Ajay for your suggestions.
I could culture the add the cells to the surface of hydrogels and they proliferated well inside the hydrogel after growing in the 3D environment
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