摘要
目的探索获得瘢痕疙瘩来源的高纯度间充质干细胞(MSCs)的培养方法。方法来源于第四军医大学唐都医院皮肤科住院的4例患者的瘢痕疙瘩标本,所取病变部位均位于前胸及后背皮肤,采用DMEM∶F12/(100mL/L)胎牛血清(FBS)初步培养,再改用低糖DMEM/(100mL/L)FBS培养,接着用低糖DMEM/(10mL/L)FBS培养,最后用无血清的干细胞培养基(SCM)培养,并通过观察细胞形态特征及应用流式细胞术检测细胞表面标记物的表达情况进行鉴定。结果经血清梯度培养后得到了长梭形、形态均一,纯度较高的纤维样细胞,经流式细胞仪检测后细胞表面标记物表达情况为:CD29为99.99%,CD44为99.7%,CD45为0.69%,CD73为99.96%。结论血清浓度逐渐降低的梯度培养法可获得高纯度的瘢痕疙瘩MSCs。
Objective To investigate the method to culture high-purity keloid-derived mesenchymal stem cells （MSCs）. Methods The keloid tissue samples were collected from the skin of 4 patients hospitalized in our department. The lesioned areas were the chest and the back. The human keloid cells were cultured with the serial culture medium in the following order： DMEM： F12/（100 mL/L）fetal bovine serum （FBS）, then low-glucose DMEM/（100 mL/L）FBS, low glucose DMEM/（10 mL/L）FBS, and last serum-free stem cell medium. Then we observed cell morphology and identified cell surface markers by flow cytometry method. Results We obtained high-purity fiber-like cells of long spindle and uniform shape by the serum gradient culture method. The expression of cell surface markers detected by flow cytometry was 99.99% for CD29, 99.7% for CD44, 0.69% for CD45, and 99.96% for CD73. Conclusion High-purity keloid-derived mesenchymal stem cells can be obtained by the culture method of decreasing serum concentration gradient.
出处
《西安交通大学学报（医学版）》
CAS
CSCD
北大核心
2014年第6期855-858,共4页
Journal of Xi’an Jiaotong University（Medical Sciences）
基金
国家自然科学基金资助项目(No.30900772)~~
