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作 者:杜江榕[1] 张献共[1] 王丹华[1] 李继云[1]
机构地区:[1]北京大学深圳医院脊柱外科,广东深圳518036
出 处:《汕头大学医学院学报》2007年第2期72-74,共3页Journal of Shantou University Medical College
摘 要:目的:探讨人脐血间充质干细胞(MSC)的培养纯化条件。方法:比较不同的梯度离心速度及时间、接种密度、首次换液时间、不同培养基对脐血中的MSC分离及原代培养过程的影响,以流式细胞仪对优化条件下培养的MSC进行细胞周期分析,并对细胞表面标志进行检测。结果:在其他条件不变的情况下,以1 000g×15 min的梯度离心速度及时间分离脐血为最宜,5×106/mL是脐血MSC培养的适宜接种密度,首次换液时间7 d。优化条件下培养的MSC传至3代时,几乎全部强表达CD29、CD44。结论:建立了一种优化的人脐血MSC分离纯化方法。Objective: To study the culture conditions of mesenchymal stem cells(MSC)from human umbilical cord blood. Methods: The gradient centrifuge speed and time, planting density, first medium changing and culture medium influenced on the growth of MSCs were analyzed. MSCs were identified using the surface marker by flow eytometry. Results: 1 000 g × 15 minutes(gradient gentrifuging speed and time)was the best way. 5 × 10^6/mL was the best planting density and the best time of the first medium changing was on the seventh day of the primary culture. Under optimized conditions, almost every MSC could express CD29, CD44 strongly when to the 3rd generation. Conclusion: A good method for isolation and purification of MSC from human umbilical cord blood is set up.
分 类 号:R744[医药卫生—神经病学与精神病学] R615[医药卫生—临床医学]
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