A Preliminary Study On Vitrification Preservation Of ASCs/demineralized Bone Tissue Complex

Objective:adipose-derived stem cells(ASCs)have become the main cell source for tissue-engineered bone.There is of great significance for bone tissue engineering to cryopreserved ASCs/scoffolds.At present,vitrification preservation is the most promising methods of cryopreservation for cells/scoffolds.Thus,to develop a novel and effective vitrification fluid mainly composed of trehalose and dimethyl sulfoxide,will be very important for cells/biomaterials complex preservation.The effect of vitrification method for ASCs/scoffolds was wished to enhance by improving the vitrified solution and the pretreatment conditionMethods:Engineered bone complex was firstly fabricated by inoculating the isolated ASCs onto the demineralized bone,followed by 7 days osteogenic inducation.Using the different composition and concentration of vitrification fluid,the freezing experiments on osteogenically induced engineered bone complex were performed under different precondition.Seven days later,cryopreserved engineered bone complex was thawed.According to the cellular survival rate,the gene expression of ALP and OCN at the third day after rewarming,the ideal vitrification media and optimal pretreatments were chosen to cryopreserve the cells using multiple response optimization AnalysisResults:The optimal composition of the novel vitrification was DMSO(v/v)30%,LG-DMEM(v/v)70%,trehalose 0.8M.Conclusions:Based on these results,it can be concluded that ASCs/scoffolds frozen by vitrification with the novel DMSO/trehalose/LG-DMEM vitrification fluid can be used to provide the massive ready-to-use cells/scoffolds for bone tissue engineering.