摘要
目的 探索成人脂肪组织源性间充质干细胞 (ADMSCs)的分离、体外培养 ,为其将来应用于临床心肌梗死的治疗提供实验依据。方法 无菌条件下获取腹部手术病人大网膜脂肪组织 ,胰酶消化 30min,离心 ,在含 1 5 %胎牛血清的DMEM中 ,37℃、5 %CO2 饱和湿度下常规培养 ,对第 2代细胞进行免疫组织化学染色 ,鉴定其表面分子CD4 4、HLA DR及Ⅷ因子。结果 成人脂肪组织中含有大量间充质干细胞 ,免疫化学染色鉴定除CD4 4阳性间充质干细胞外 ,所分离细胞同时含有部分HLA DR阳性成纤维细胞和Ⅷ因子阳性的血管内皮细胞。结论 建立了一种自人体脂肪组织分离、培养ADMSCs经济简便的方法 。
Objective To look for a method for isolation and cultivation of mesenchymal stem cells from human adipose tissue in vitro. Method The adipose tissue was obtained from the omentum of abdominal surgery patients under the aseptic condition. The fat then was minced and digested with 5ml 0 25% trypin for 30 minutes in a 37℃ water bath under constant agitation. The layer with monoclear cells was aspirated and supplemented with albumin, then was cultured in DMEM with 15% calf serum. Immunocytochemistry was used to determine the surface molecule CD44, HLA DR and VWF. Result There was a large amount of mesenchymal stem cells in human adipose tissue. Immunocytochemical staining showed that most of the cultured cells were CD44 positive, few HLA DR positive and VWF positive, indicating that most of the cultured cells were MSC, and the others were fibroblasts and endothelial cells. Conclusion A simple and convenient method to isolate and culture the adipose tissue derived mesenchymal stem cells was successfully established, providing the foundation for the future use of ADMSCs in the treatment of cardiovascular diseases.
出处
《解放军医学杂志》
CAS
CSCD
北大核心
2004年第6期510-512,共3页
Medical Journal of Chinese People's Liberation Army
基金
国家自然科学基金资助课题 (编号C30 0 0 0 1 96)
关键词
间充质干细胞
脂肪组织
细胞培养
mesenchymal stem cells
adipose tissue
cell culture
