Serum-free cell cryopreservation based on self-assembled peptides
Objective To achieve serum-free and low-DMSO efficient cell cryopreservation based on self-assembled pep-tides(Peptide).Methods Hepa1-6 cells were cryopreserved by combining three cryoprotectants(CPAs),FBS,DMSO and Peptide.Confocal microscopy,flow cytometry and cell counting were used to measure the survival rate,recovery rate and proliferation ability of the cells after freezing,and the ice inhibition of CPAs was tested by differential scanning calori-metry(DSC).Finally,a CPA with high efficiency for cell cryopreservation was determined.Results Peptide formed a fibrous network hydrogel in DMSO solution to encapsulate cells and supported them in a three-dimensional(3D)stereological distri-bution.Under serum-free conditions,the use of 0.1 wt％Peptide combined with 5％DMSO as a novel composite CPA for cryopreservation of Hepa1-6 cells significantly improved the survival rate and post-freezing recovery rate compared with the 5％DMSO and 0.1 wt％Peptide groups,and there was no significant difference with the 10％DMSO+20％FBS group(t=0.983 and 2.164,respectively,each P＞0.05).In addition,compared with the 10％DMSO+20％FBS group,0.1 wt％Peptide+5％DMSO group showed no significant difference in cell proliferation ability after freezing-thawing(t=3.513,P＞0.05).In the end,DSC tests showed that Peptide-based CPA had good ice inhibition performance.Conclusion The use of 0.1 wt％Peptide combined with 5％DMSO as a novel composite CPA has a good effect on cell cryopreservation.
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