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OALib Journal期刊
ISSN: 2333-9721
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双阴性选择法纯化培养人骨髓多能成体祖细胞
, PP. 535-537
Keywords: 人骨髓来源多能成体祖细胞,免疫磁珠分离法,分离纯化,细胞培养,流式细胞术
Abstract:
目的建立体外分选纯化及培养人骨髓来源多能成体祖细胞(mapcs)的方法.方法取健康成人志愿者适量骨髓采用梯度密度离心法分离获取单个核细胞,贴壁培养后,将获取的骨髓贴壁细胞通过cd45、血糖蛋白a(glya)免疫微磁珠(minimacs)负分选,流式细胞术鉴定分选后细胞纯度;对传代到不同阶段的细胞进行cd45、glya流式细胞分析,初步了解mapcs在培养、增殖过程中的免疫学稳定性.结果通过minimacs分选,平均每1×106/ml骨髓贴壁细胞可分选出约(5～10)×104/ml的mapcs,分选前后细胞活力分别为(96.7±1.7)%和(96.0±2.4)%,无明显差异;自制的培养基培养分选后的mapcs生长良好,最长传代到第16代;流式细胞仪分析获取的cd45、g1ya-细胞纯度大于98%;流式细胞仪分析传代第4、8、12代mapcs,cd45-、g1ya-细胞纯度大于98%.结论cd45、glyaminimacs负分选可从骨髓中分离高纯度的mapcs;mapcs在本室自制的人mapcs培养基中体外有较强的增殖能力,且能保持较长时间的稳定状态.
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