I have found that my cells, NIH3T3 fibroblasts, do not do great on gelatin. The number of cells that adhere or spread after 24 hours and even further are far less on gelatin (5 and 10%) than on chitosan (2 and 4 %). The expected outcome is generally the opposite. All the gel samples were given same treatment.
Any suggestion is appreciated!
It can't be unless you autoclaved gelatin sol, or you coated gelatin sol properly on cleaned glass coverslips by standing gelatin drop on the coverslip for 30 min, dried for 1h, washed with ddH2O, dried again. I found my students get this problems with an easy prep. by soaking coverslips for a brief time or if gelatin is dissolved in salt sol.
1. Check your 3T3 cell are clean by staining with DAPI to check any bac or mycoplasma.
2. Check your medium whether it is sterile.
3. Prepare dust-free, clean coverslip, dried
4. Make sure your gelatin sol is new and clean, if you have any doubt or left from someone else , spin the sol in 1ml eppen tube at 12,000rpm for 10 min to remove any debris or contaminated microbes. Use the supernatant.
5. Put 50ul of gelatin sol on the middle of a small round coverslips, and leave them to dry completely under dust-free lamina flow within the 100ml culture dish. (give 1h)
6. Wash the coverslips in ddH2O (the stacked gelatin molecules loose should be washed away leaving a nice coating layer of gellatin molecules) and dry them and store in glass petri dish with lid.
Hi!
Seed at least 3 walls on collagen covered plastic and compare. It could be a lot of explanations. May be, gelatine include any compound used for its chemical processing (for example, any acid).
what did you use to crosslink gelatin scaffold?
and did you dissolve gelatin in d.water or in acetic acid?
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