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体外培养单个造血干细胞实验方法的建立 被引量：3

Establishment of experimental methods to culture single hematopoietic stem cell in vitro
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摘要 目的通过建立一种体外稳定培养单个造血干细胞(hematopoietic stem cell,HSC)的实验方法,在未来筛选可以促进HSC体外扩增的化合物。方法通过配制HSCs体外培养液并结合流式分选技术,我们建立了一种在体外不需要基质细胞支持的,稳定的,高通量培养单个HSC形成一个血细胞集落的实验方法。结果只需培养10～14 d即可观察到化合物对HSC的作用,并结合流式细胞分析技术替代人工计数血细胞分化情况,进一步节省人力,具有客观、重复性强的特点,满足了高通量筛选药物的要求,为后续的研究和化合物筛选工作打下基础。为了验证此方法的可靠性,使用BIO(一种通过抑制糖原合成酶激酶-3,进而可以促进HSC自我更新的化合物)时,实验结果与文献报道的研究结果一致。结论该实验方法在有关造血干细胞体外扩增的药物筛选方面具有很好的应用价值。 Aim To establish a method to stably culture a single HSC in vitro, to screen the compounds which can promote the self-renew of hematopoietic stem cells （HSCs）. Methods By the configuration of HSCs in vitro culture medium and flow-cytometry sorting, a method was established to stably and highthroughput culture a single HSCs without the stroma cell support. Results The compound effect on the HSC was found inonly 10 to 14 days. The flow analysis of the differentiation, which substitutes the manual count, is a future saving of manpower. Conclusions This method is objective , reproducible and can be used in the high throughput screening of drugs.
作者 李乔 纪庆 王金宏 许静 田晨 程辉 刘延风 张娜 程涛 高瀛岱
机构地区 中国医学科学院血液病医院(血液学研究所)实验血液学国家重点实验室
出处 《中国药理学通报》 CAS CSCD 北大核心 2012年第3期435-438,共4页 Chinese Pharmacological Bulletin
基金 国家自然科学基金资助项目(No 81170465 90913018 81090411) 实验血液学国家重点实验室开放课题基金(ZK11-07 ZK08-04)
关键词 单个HSC培养 流式细胞分选与分析 血细胞涂片 P18 小分子 干细胞扩增 single hematopoietic stem cell in vitro culture flow cytometry sorting and analysis blood smear p18 small molecule HSC proliferation
分类号 R329.24 [医药卫生—人体解剖和组织胚胎学] R329.28 [医药卫生—人体解剖和组织胚胎学]
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参考文献13

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二级参考文献17

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同被引文献33

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二级引证文献16

中国药理学通报
2012年 第3期

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