I have synthesized cryogels to perform the in vitro biochemical assay, which requires sterilized cryogels as I am seeding cells onto it. Therefore, I am looking for a standard protocol for the sterilization of cryogels. Thank you.
Hi Isha Behere, You can dip the cryogel in 70% ethanol with or without UV irradiation and be given time to ensure complete penetration. Afterward, ethanol needs to be removed aseptically by dipping cryogel in sterile PBS and media before proceeding to cell culture.
Hello, Isha Behere
Having considered the information on cryogels sterilization, in our lab now we use the storage in 70% ethanol as the easiest and most accessible way to do it. Before cell culturing, we wash cryogels in sterile water twice, and once in cell culture medium. When replacing the liquid, the cryogels are pressed to get rid of ethanol. Still there are some other methods listed in literature. One of them is sterilizing using ultraviolet light in a laminar flow chamber for 30 min [Carvalho DN, López-Cebral R, Sousa RO, et al. Marine collagen-chitosan-fucoidan cryogels as cell-laden biocomposites envisaging tissue engineering. Biomed Mater. 2020;15(5):055030]. Another is washing several times with sterile PBS and then sterilizing with UV for 1 h [Kim HD, Kim J, Koh RH, et al. Enhanced Osteogenic Commitment of Human Mesenchymal Stem Cells on Polyethylene Glycol-Based Cryogel with Graphene Oxide Substrate. ACS Biomater Sci Eng. 2017;3(10):2470-2479]. The third one is sterilizing by gradient ethanol (20%, 40%, 60%, 80%, 100%) followed by three times PBS wash of 15 min each, after PBS wash cryogels discs are incubated in complete medium overnight at 37 °C in a CO2 incubator [Kumari J, Kumar A. Development of polymer based cryogel matrix for transportation and storage of mammalian cells. Sci Rep. 2017;7:41551].
Hope it has been helpful.
Thank you Jijo Thomas and Yu. B. Basok for the valuable information. I would also like to know, does UV light affect the structural properties of cryogels? Thank you once again!
Hi Isha Behere , Normally UV don't affect structural properties. But it may vary on the response of the polymer used to the UV light.
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