Our lab has been working to reduce DMSO in our media for hepatocyte cryopreservation. Do you have any recommendations on reducing DMSO along with the lowest percentages you’ve seen for this application.
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The Dos and Don’ts of Cryopreservation
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There are many reports regarding cryopreservation of hepatocytes from different species like mouse, monkey, dog, rat, and human. According to many literature, the authors used 10-20% of DMSO for cryopreservation of hepatocytes. 10% of DMSO is most common concentration as a minimum of cryoprotectant for hepatocytes. If you are worried about cell viability, you can attempt to add oligosaccharides as a supplement in the freezing media that contains 10% of DMSO(1). The authors observed use of oligosaccharides resulted in greatest improvement in cell viability using trypan blue (TB) exclusion method. In addition, a researcher team in Sweden evaluated cell viability of hepatocytes using STEM-CELLBANKERTM (CB) which is a xeno-free cryopreservation solution containing 10% DMSO, glucose and anhydrous dextrose, comparing standard 12% DMSO-UW medium(2). They observed higher cell viability in CB protocols than DMSO-UW in hepatocytes.
(1) https://journals.sagepub.com/doi/pdf/10.3727/000000006783981404
(2) https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3365437/
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