I am trying to sterilise high molecular weight polyethylene oxide (~400,000 Da) in DMEM for culturing bmMSCs. I have done online search for relevant information, but most studies were using PEG less than 4000Da. I am comparing the effect of polyethylene oxide at 400,000 Da, with another substance of similar molecular weight, in mimicking tissue microenvironment to promote extracellular matrix deposition.
I run into problem of sterilising the solution. I am currently trying to prepare a maximum concentration of 25mg/ml. I have tried filtering with 0.22um and 0.45um filters, but it does not work.
Data sheet of PEG 8000 from Sigma indicated that solution can be prepared with 0.45um filters, but it was not stated at what concentration, and the molecular weight is different after all. I was not able to filter with 0.45um filters anyway. It also indicate that autoclaving is not recommended, even mentioning that while many people do it, they do not recommend it.
Then I was thinking of sterilisation under UV, but I also read that prolong UV exposure (1 hour) can resulting in degradation. However, again these studies were done with PEG of a different molecular weight.
I am wondering if anyone have experience or suggestion on how I can sterilise the substance. Any opinion will be greatly appreciated.
Dear Kwok Keung Lit, high MW PEO's have somehow similarities to crosslinked PEG hydrogels. The following document deals with sterization pecularities of PEG's hydrogels. My Regards
https://doi.org/10.1002/jbm.a.31811
Dear Kwok Keung Lit, high MW PEO's have somehow similarities to crosslinked PEG hydrogels. The following document deals with sterization pecularities of PEG's hydrogels. My Regards
https://doi.org/10.1002/jbm.a.31811
Hi;
PEO can be sterilized chemically by immersing in isopropanol/H2O mixture, 70/30% (v/v) or in ethanol/H2O mixture, 70/30% (v/v) followed by rinsing with phosphate buffered saline (PBS).
With my best regards
Kindly check https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6591268/
Also check https://www.researchgate.net/publication/338957917_Polystyrene-_block_-polyethylene_oxide_thin_films_In_vitro_cytocompatibility_and_protein_adsorption_testing
Thank you all for your suggestion. Although none has been applicable for my application, I appreciated for all your suggestions. They are helpful in eliminating some methods, as suggested by the reference provided by Abdelkader BOUAZIZ.
I am using bone marrow messenchymal stem cells (which is adherent cells), and I am not fabricating the PEO into film. The PEO we had purchased is in powder form.
I am now thinking of dissolving them in 70% ethanol, and freeze drying them before dissolving them again in the DMEM for use. Will update if it works.
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