Dear all,
I am culturing human primary keratinocytes and fibroblast together to make an organotypic model of skin. I already had the following collagen solution in my lab so I used it.
https://www.sigmaaldrich.com/catalog/product/sigma/c3867
Unfortunately, this solution does not transform into a 3D gel when I add NaOH in it.
If anyone has a similar experience then kindly guide me.
Or,
is there any possibility that these cells would make a differentiated skin model without a solid collagen layer below them?
Have you tried filter culture? https://www.thermofisher.com/us/en/home/life-science/cell-culture/cell-culture-plastics/cell-culture-inserts/cell-culture-inserts-skin-tissue-culture.html
I have done extensive work on organtypical culture using primary and cell line oral keratinocytes, you find more detail on my paper:
Article Tissue engineering of oral dysplasia
I hope this may help you
Perhaps you should try to replace the type of collagen in your solution, or add another substance capable of doing the cross-linking reaction (fibronectin, laminin, gelatin).
I'm also working with 3D collagen solutions for cell coating processes, and in my case, I have been associating the Collagen type IV (which corresponds to the negative charge) with laminin (which corresponds to the positive charge). Another possibility, perhaps, would be to check some commercially available ECM-based mixture, such as Matrigel.
Take a look at these links, maybe it would help you.
>https://www.sigmaaldrich.com/catalog/product/sigma/c6745?lang=pt®ion=BR&cm_sp=Insite-_-caRecBlock_recBlock_cooccuranceModel-_-recBlock5-3
> https://www.sigmaaldrich.com/catalog/product/sigma/e1270?lang=pt®ion=BR
> https://www.sigmaaldrich.com/catalog/papers/25689062
> https://www.sigmaaldrich.com/catalog/product/sigma/l2020?lang=pt®ion=BR
Good luck!
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