Hi everyone, I am using atecollagen from Koken company to make the cell beads for 3D cell culture. Intially, pH of the gel is 7.3-7.5, which is suitiable for gelation. However, After put these gel in 37oC incubator for 1hour, it cannot happend the gellation, so the stiff is not enough to maintain the bead shape. Can you guys help me how to improve it? Thank you
What is the collagen concentration in the stock solution and what is the final concentration after preparation? Possibly the collagen is too dilute to form a strong matrix.
Thank you, Matthias Gossmann. Both of them are 2 mg/ml and no diltution here. Actually, I just want to use only thermal-crosslinking for gelation without any chemical-crosslinking reagents. but maybe it is not enough.
You might try a pre-incubation at 4C and slightly lower pH. See Sung, et al., Biomaterials. 2009 Sep; 30(27): 4833–4841. for additional information. Also Google collagen gelation cold temperature for additional references.
I suspect that "Atecollagen" is collagen that had its telopeptides removed (by enzymatic treatment). I thin that such collagen do not form strong gels. Read carefully the description of the product and if that is the case, buy acid-solubilized collagen.
Hi,
You may try to dilute it into 1.0-1.5mg/ml solution with 10% FBS DMEM.
I think Nicolas' suspicion may be right, so you should neutralize the protease by FBS and other salt in the medium may help gelation.
And please let me know the result if you try it.
Thank you, everyone for your advices,
After changing the atelocollagen into acid-solubilized native collagen, The mechanical properties increased and collagen structure was mantained, fortunately .
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