Dear all,
I am trying to culture human neonatal fibroblast with a keratinocytes together to make an organotypic skin model.
Currently, I am working according to the established protocol: https://pubmed.ncbi.nlm.nih.gov/17401352/
The difference is that I had already used a created collagen solution: https://www.sigmaaldrich.com/catalog/product/sigma/c3867
But the solution does not transform into a 3D gel, even if I add 1M NaOH in it, or by adjusting the pH of the solution to a neutral range.
Is there any other problem, e.g. in collagen, temperature, ...?
Thank you for your answer!
Replace the kind of collagen in your solution if possible, or add another component capable of causing the cross-linking process (fibronectin, laminin, gelatin).
I'm also experimenting with 3D collagen solutions for cell coating procedures, and I've been coupling Collagen IV (which has a negative charge) with laminin in my instance (which corresponds to the positive charge). Another option would be to look at a commercially available ECM-based mixture, such as Matrigel.
Take a look at these sites to see if they may assist you.
You can solve your problem below the link:
https://www.sigmaaldrich.com/BD/en/product/sigma/c6745?cm_sp=Insite-_-caRecBlock_recBlock_cooccuranceModel-_-recBlock5-3
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