Effect of adipose tissue storage conditions on in vitro isolation and culture of human adipose mesenchymal stem cells
Objective To investigate the effects of antibiotics,adipose tissue preservation time and tem-perature,culture medium on in vitro isolation and primary culture of human adipose mesenchymal stem cells(ADMSC).Methods The superficial abdominal subcutaneous adipose tissues collected by liposuction were stored at 2-8 ℃,room temperature(22 ℃),37 ℃ for 0,1,2,3,5,7 d.After that,the cells were extracted by the enzyme digestion method for conducting the primary culture,purified by the cell adherent method,diges-ted and passed by the pancreatic enzyme.Phenotypes were detected by flow cytometry in P5 generation ADM-SC.The P5 generation ADMSC were induced to differentiate in adipogenic and osteogenic induction solution,and identified by Oil Red O staining and alizarin red staining.Results The primary ADMSC isolated from adi-pose tissue without gentamicin passed 3 d earlier than those isolated from adipose tissue with gentamicin.The first passage day of primary ADMSC isolated from adipose tissue stored for 0,1,2,3,5 d was 5,6,6,7,9 d,re-spectively,and the primary ADMSC isolated from adipose tissue stored for 7 d could not be amplified normal-ly.The average cell yield of primary cells isolated from adipose tissue stored at 2-8 ℃,room temperature and 37 ℃ on 6 d was(7.7-8.4)×104/cm2,(6.9-7.4)× 104/cm2,(4.8-5.2)×104/cm2,respectively.Fat in the medium containing nucleoside was transited 2 d earlier than fat in the medium without nucleoside.After in-duction of adipogenesis,the induced cells exhibited the characteristics of adipocytes and osteoblasts,respec-tively,and which in the control group was all negative.Conclusion Antibiotics,long-term,high temperature storage inhibit ADMSC proliferation,while nucleosides could promote its proliferation.
adipose mesenchymal stem cellsisolation and culture in vitrostorage temperaturestorage timeantibioticsculture medium
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