重庆医学2024，Vol.53Issue(7)：972-978,7.DOI:10.3969/j.issn.1671-8348.2024.07.003
脂肪组织保存条件对体外分离培养人脂肪间充质干细胞的影响
Effect of adipose tissue storage conditions on in vitro isolation and culture of human adipose mesenchymal stem cells
摘要
Abstract
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.关键词
脂肪间充质干细胞/体外分离培养/保存温度/保存时间/抗生素/培养基Key words
adipose mesenchymal stem cells/isolation and culture in vitro/storage temperature/storage time/antibiotics/culture medium分类
医药卫生引用本文复制引用
梁坤荣,薛冰华,王慧,顾恩妍,吴振奇,石峰,靳丽艳,张蕊,于婷婷,王艳超..脂肪组织保存条件对体外分离培养人脂肪间充质干细胞的影响[J].重庆医学,2024,53(7):972-978,7.基金项目
国家重点研发计划项目(2020YFA0112803) （2020YFA0112803）
北京市顺义区重大科技研发项目(2022). （2022）
