I have tried so many techniques to keep my cells viable in keratin gels. Unfortunately, cells dying after 24 hours.
Can you please elaborate your protocol in simple and short sentences so that we may understand better whats happening? And whats causing your samples to be toxic.
Hello Neda,
Can you please describe how you extracted Keratin? there are numerous techniques and in many cases, the final product is toxic to cells. e.g. using mercaptoethanol, urea and sulphur salts can have residues and make keratin toxic.
Would I suggest a good dialysis
in addition have you checked the molecular weight of your keratin? high MW oligopeptides are not cell friendly either
Please see this paper, hope it helps
Thank you Anwaar, Amin, and Mamatha for your willingness to help me. Keratin was provided by one of our collaborators. The procedure is summarized in the attached paper. As it is mentioned, keratin was dialyzed for 24 before lypholization step.
I have got the keratin powder and dissolved in media for the final concentration of 28 mg/ml. The solution was mixed with cells and added to the 98 well plates. So, cells are embedded in the matrices. Cell viability was measured over the course of seven days; however, after 24 hours 90% of cells were dead.
I appreciate any comments you have. Thanks much in advance :)
Hello Neda,
Since the keratin was dialyzed, I would recommend lowering the concentration of keratin (between 10 to 15 mg/mL). It should be more than enough to form gel/hydrogel.
Hi Lukasz,
We have tried 3 mg/ml keratin as well. But got the same outcome! Can it be cells attachment issue?
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