Hello,
I am using Corning Rate Tail collagen Type I for the synthesis of skin tissue scaffold. I am using 2.5 mg/mL of collagen, 10 X DMEM, Fibroblasts cells and FBS. All the steps were performed on ice and pH was adjusted to 7.4 using 0.5 M NaOH.
Even this is standard protocol in most of the references, I am unable to get collagen in gel form. It remains in liquid form. Can anyone suggest me the correct protocol for collagen gelation and skin scaffold preparation.
Of some possible interest for your query: https://www.researchgate.net/post/How_to_make_my_rat_tail_collagen_I_gel https://www.researchgate.net/post/What_is_the_proper_collagen_gel_preparation_method2
Hi,
You need to incubate the collagen solution mixed with fibroblast in 37 0C to gel. It works for me all the time. It can also be gelled at 4 deg for a longer time but the fiber would be thicker. Make sure the pH is neutral or basic to allow it to gel.
I use the protocol from Corning.
Also, make sure the collagen is not expired and stored at 4 deg.
Sunil
Thanks everyone for addressing my query.
for this experiment I used newly purchased collagen from corning (3.75 mg/mL)
Protocol used is as follow
900 µL of Collagen solution (in 0.02 M Acetic acid as directed by manufacturer)
35 µL of 10 X PBS with phenol red.
pH was adjusted with 1 M NaOH (~15 uL)
60 µL of FBS
100 µL of 10 X DMEM
50 µL of cell suspension
Incubated at 37°C in CO2 incubator.
Shailaja Agrawal
I am facing the same problem. I adjusted the pH by using pH paper. Even after 2/3 hrs of incubation, I did not get the gel. The solution became cloudy but no gel form. Did you solve your problem? if so would you please share with me?
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