I would like to recover live cells from a hydrogel containing gelatin, sodium alginate and sodium chloride. After introduction of the cells the hydrogel was crosslinked with calcium chloride.
I heard that the mixture of EDTA and sodium citrate is perfect for dissolving such hydrogels. So I tried it in different concentrations (0.1M-1M), temperature (15-40), pH (5-9).
Unfortunately, nothing happened even after overnight treatment. Now, I was wondering whether somebody had the same problem and how did you manage to resolve it.
Thank you in advance.
Though I've never had experience on such mixture, I think the gelatin remains and keeps the structure, because sodium alginate gel is normally dissolved by sodium citrate readily. Did you try to disolve gelatin using trypsin or so?
No, I have not used trypsin.
What kind of trypsin would you recommend for the digestion?
I found this on Sigma: http://www.sigmaaldrich.com/catalog/product/sigma/t4549?lang=en®ion=GB.
As this experiment is done in London that would be quite convenient.
http://www.sigmaaldrich.com/catalog/product/sigma/t4549?lang=en®ion=GB
I think the trypsin in your link works.
FYI: Trypsin with EDTA is also available
http://www.sigmaaldrich.com/catalog/product/sigma/t4174?lang=en®ion=JP&cm_sp=Insite-_-prodRecCold_xviews-_-prodRecCold10-9
The solvent is not indicated. I suppose the question is related to water. They are not indicated concentrations.
However, the gelatin must be dissolved in hot water. The same applies for the sodium alginate and the two solutions should be combined, by mixing hot.
It is not possible to dissolve the calcium chloride together with the sodium alginate because they form the calcium alginate, insoluble in water, which precipitate.
It is possible add the calcium ions by immersing the gelatin gel containing sodium alginate, at room temperature, in a solution of calcium chloride in water.
It will form the interpenetrating hydrogel of calcium alginate and gelatin.
As for the last part of the question (EDTA and sodium citrate) I do not answer because I have never worked with these compounds.
The preparation of the hydrogel has been done as L.A. Pajewski described.
It seems the problem was the calcium chloride concentration.
A soon as we decreased that we succeeded with the dissolution using EDTA and sodium citrate.
I think we will try the trypsin as we still have the gelatin. Thank you Nobuhito Mori.
Thanks both of you for your assistance.
Csaba Sarosi , I was wondering if you managed to solve your problem and dissolve the hydrogel. If so, would you be willing to share the method you used to do so?
The following discussions seemingly can be found of some interest, concerning to your query: https://www.researchgate.net/post/Does_change_in_pH_of_sodium_alginate_affect_final_encapsulated_cell_viability https://www.researchgate.net/post/How_can_I_make_alginate_disks_that_I_can_put_into_a_24-well_plate/1
About predicting the pH of CaCl2 aq. solutions ― cf. my posts at: https://www.researchgate.net/post/How-to-increase-pH-for-a-solution-without-making-a-chemical-reaction-with-CaCl2
About predicting the pH of citric acid ― sodium citrate solutions and buffers for approx. pH < 4.0 ― cf. my post at:
https://www.researchgate.net/post/How-to-prepare-a-citric-acid-solution-of-pH-25-and-3-from-100-citric-acid-powder
About predicting the pH of citric acid ― sodium citrate solutions and buffers for approx. 4.0 < pH < 5.5 ― cf. my posts at:
https://www.researchgate.net/post/Why-the-pH-of-Citrate-buffer-increases-when-diluted-with-water
About predicting the pH of citric acid ― sodium citrate solutions and buffers for approx. 5.5 < pH < 8.0 ― cf. my posts at: https://www.researchgate.net/post/Why-the-pH-of-Citrate-buffer-increases-when-diluted-with-water
About predicting the pH of citric acid ― sodium citrate solutions and buffers for approx. pH > 8.0 ― and for approx. pH > 9.0 ― cf. my posts at: https://www.researchgate.net/post/Why-the-pH-of-Citrate-buffer-increases-when-diluted-with-water
Hoping you have sort out your issue. I would recommend 50:50 water glycerol and treat it will ultrasounic homogenizor of 3 MM horn for 20 min
At 300W of 3 sec puls
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