人脐带血间充质干细胞的分离培养及增殖能力

背景:随年龄的增加,骨髓来源的间充质干细胞在数量和质量上均受到影响,并对供体损伤较大,寻找替代的间充质干细胞来源成为研究热点,而脐带血中是否含有间充质干细胞仍存在争议.目的:拟从人脐带血中分离培养间充质干细胞,并对其生物学特性进行检测.方法:无菌条件下,应用Percoll密度梯度离心法分离脐血标本,收获中间层细胞,加入含胎牛血清、青霉素和链霉索、L-谷氨酰胺的DMEM基础培养液,再经反复贴壁纯化,除去悬浮生长细胞,得到较多呈融合状态的贴擘细胞,待细胞达60%～80%融合时胰酶消化传代.分别取第1,5,9代细胞,观察细胞形态变化,流式细胞仪检测细胞表面抗原的表达,绘制细胞生长曲线,MTT法检测细胞活性.结果与结论:分离的脐血间充质干细胞大小均匀,呈梭形或星形的成纤维细胞样细胞.第1,5,9代脐带血间充质干细胞高表达CD29,CD105和CD166,低表达CD34和CD45:接种后5 d细胞进入指数增生期,9 d后数量减少,群体倍增时间为(53.5±8.32)h;细胞生长状态良好,在细胞活力方面1～7代摹本相似(P＞0.05),至第9代细胞增殖活力稍下降,但仍无明显差异(P＞0.05).结果证实可在体外成功从脐血中分离培养出间充质干细胞,第1～9代细胞均具有良好的增殖能力.

Isolation,cultivation and reproductive activity of human umbilical cord blood mesenchymal stem cells

BACKGROUND: With increased age, bone marrow mesenchymal stem cells (BMSCs) are influenced with regard to quantity and quality, which will induce great damages to the donors. Many studies have focused on seeking substitute MSC source. In contrast, it remains controversial whether umbilical cord blood contains MSCs. OBJECTIVE: To isolate MSCs from human umbilical cord blood, and to detect their biological properties. METHODS: Umbilical cord blood samples were sterilely isolated using Percoll density gradient ...