检索规则说明:AND代表“并且”;OR代表“或者”;NOT代表“不包含”;(注意必须大写,运算符两边需空一格)
检 索 范 例 :范例一: (K=图书馆学 OR K=情报学) AND A=范并思 范例二:J=计算机应用与软件 AND (U=C++ OR U=Basic) NOT M=Visual
作 者:丁丽[1] 朱恒[2] 张海宏[3] 杨洋[1] 韩冬梅[1] 王志东[1] 郑晓丽[1] 董磊[1] 闫洪敏[1] 刘静[1] 朱玲[1] 薛梅[1] 郭子宽[4] 王恒湘[1] DING Li ZHU Heng ZHANG Hai-hong YANG Yang HAN Dong-mei WANG Zhi-dong ZHENG Xiao-li DONG Lei YAN Hong-min LIU Jing ZHU Ling XUE Mei GUO Zi-kuann WANG Heng-xiangl(Department of Hematology, Air Force General Hospital of Chinese PLA, Beijing, 100142, China Institute of Basic Medical Sciences, Academy of Military Medical Sciences, Beijing, 100850 Department of General Surgery, Air Force General Hospital of Chinese PLA, Beijing, 100142 Institute of Radiation Medicine, Academy of Military Medical Sciences, Beijing, 100850)
机构地区:[1]中国人民解放军空军总医院血液科,北京100142 [2]军事医学科学院基础医学研究所,北京100850 [3]中国人民解放军空军总医院普外科,北京100142 [4]军事医学科学院放射与辐射医学研究所,北京100850
出 处:《中国比较医学杂志》2016年第10期56-60,共5页Chinese Journal of Comparative Medicine
基 金:国家自然科学基金项目(81500083;81371945;81572159;81101342);空军总医院院级课题(KZ2014023);北京市自然科学基金项目(7132133)
摘 要:目的通过组织块培养法,建立一种高效可靠的小鼠脾脏间充质干细胞分离扩增策略。方法无菌条件下取小鼠脾脏组织充分碾碎,使用胶原酶消化获得的脾脏基质组织,将消化后的组织块种于培养瓶中,观察从组织块中迁移出的细胞形态,采用流式细胞术分析其表型,并将其做成骨和成脂肪诱导分化。结果消化后的脾脏组织块中迁移迁移出大量梭形纤维样细胞,流式细胞术检测结果表明,该种细胞高表达CD29、CD44、Sca-1和CD105,低表达CD11b、CD34、CD45和Ia。多向分化实验结果表明:小鼠脾脏基质组织中迁移出的细胞具有较好的成脂肪和成骨分化能力。结论组织块法可以分离培养出小鼠脾脏间充质干细胞,所获得的细胞纯度高,分化能力强,为开展相关研究提供了良好的细胞模型。Objective This study aimed to establish a reliable primary culture protocol for preparing murine spleen-derived mesenchymal stem cells( MSCs) by tissue explant culture. Methods Healthy mouse spleens were crushed by syringe handle to harvest spleen mesenchymal tissues. Then the tiny pieces of spleen tissue were digested by collagenase II before seeded into culture flasks. The morphological characteristics of spleen tissue-derived cells were observed under the inverted microscope. Further,the surface antigen profile of the cells was analyzed by flow cytometry( FACS). The cells were induced to differentiate into osteoblasts and adipocytes. Results The murine spleen-derived MSCs exhibited a spindle-shaped appearance. The FACS results showed that the spleen-derived MSCs highly expressed CD29,CD44,CD105 and Sca-1, but weakly expressed CD11 b, CD34, CD45 and Ia. In addition, the spleen-derived MSCs steadily differentiated into osteoblasts and adipocytes in the induction medium. Conclusions A method of primary culture of murine spleen-derived MSCs by explant culture is successfully established. The harvested MSCs exhibit high purity and cell proliferation ability,and provide a reliable cell model for related researches.
分 类 号:R33[医药卫生—人体生理学]
正在载入数据...
正在载入数据...
正在载入数据...
正在载入数据...
正在载入数据...
正在载入数据...
正在载入数据...
正在链接到云南高校图书馆文献保障联盟下载...
云南高校图书馆联盟文献共享服务平台 版权所有©
您的IP:3.148.198.61