High Glucose Induces Cellular Senescence And Reduces Therapeutic Effects Of Adipose Derived-mesenchymal Stem Cells

To overcome the existing problems of separation processes for adipose derived-mesenchymal stem cells(AD-MSC),such as residual tissue block,tedious filtering steps,as well as impurity of primary cells,collagenase NB4 were used as digestive enzyme in the separation processes.The current results show that collagenase NB4 could effectively reduce the residual tissue block,simplify the digestion and filtration processes,and obtain pure P0 primary AD-MSC.Identification of surface antibody molecules indicates that the AD-MSC cells separated by NB4 are relatively pure.CD73,CD90 and CD105 are effectively expressed on the AD-MSC(?95%),while other surface antibodies such as CD14,CD34,CD19,CD 45,CD11 b,CD79a,HLA-DR or MHC II were not expressed.The separated AD-MSC cells have three-line differentiation abilities and could differentiate into fat cells,bone cells or cartilage cells,suggesting that the proposed method is capable of Improving the quality of the AD-MSC,reducing operating multifarious,and obtaining high-quality AD-MSC.In order to understand the effects of glucose concentration on the biological characteristics of AD-MSC,high glucose DMEM and low glucose DMEM culture media were used for AD-MSC.Cell morphology,immune phenotype,proliferation,senescence,cell cycle,gene expression,cytokine secretion and differentiation potential of different generations cells were detected.Results show that long-term high glucose concentration is capable of decreasing AD-MSC activity significantly,and losing the characteristics of the spindle fibroblasts gradually.In addition,the high glucose concentration could result in senescence,decreasing proliferation ability,gene expression,cytokine secretion,as well as differentiation potential.On the contrary,the low glucose environment seems no obvious adverse effects.Furthermore,the therapeutic effects of AD-MSC cells which were cultured in different glucose concentration conditions were also evaluated.The results indicate that after two doses injection of AD-MSC(cultured by high glucose media or low glucose media),the weight and fasting glucose of diabetes mouse were recovered.However,the therapeutic effects of high glucose cultured AD-MSC were much less than the low glucose cultured AD-MSC.This result further illustrates that high glucose environment may have negative influence on the therapeutic effects of AD-MSC.