High‐resolution cell division tracking demonstrates the Flt3‐ligand‐dependence of human marrow CD34+CD38− cell production in vitro
High‐resolution cell division tracking demonstrates the Flt3‐ligand‐dependence of human marrow CD34+CD38− cell production in vitro
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高分辨率细胞分裂追踪证明人骨髓 CD34+CD38− 细胞体外生产的 Flt3 配体依赖性
DOI：
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发表时间：
1997
影响因子：
6.5
通讯作者：
C. Eaves
中科院分区：
文献类型：
--
作者：
R. Nordon;S. Ginsberg;C. Eaves
原始人类造血细胞行为的调查需要监测异步激活的细胞在几代人的方法。我们描述了一种高分辨率的程序，用于跟踪5-（和6-）羧基荧光素二乙酸琥珀酰亚胺酯（CFSE）标记的人造血细胞通过6个细胞周期，基于其CFSE荧光在每个有丝分裂的精确减半。使用这种方法与DNA或表面抗原染色相结合，我们表明，向由Steel因子、IL-3、IL-6和G-CSF组成的细胞因子混合物中添加Flt 3-配体（FL）增加了CD 34+细胞的比例。（CD 45 RA/CD 71）−，但非CD 34 +（CD 45 RA/CD 71）+，最初募集到体外分裂的人骨髓细胞，缩短了其后代的总周期时间，并通过几代（最多四代）细胞增强衍生CD 34 + CD 38 −群体的产生。这些研究还表明，在前4 天，在CD 34+细胞的后代中没有可检测到的凋亡。（CD 45 RA/CD 71）-细胞在这四种细胞因子混合物的存在下产生，无论FL的存在如何。监测单个细胞特性变化的技术的可用性是其有丝分裂历史的函数，并且在它们被异步招募分裂的条件下，为研究提供了一种新的且强大的方法原始人类造血细胞增殖和分化的调节。
Investigation of primitive human haemopoietic cell behaviour requires methodologies for monitoring asynchronously activated cells over several generations. We describe a high‐resolution procedure for tracking 5‐ (and 6‐) carboxyfluorescein diacetate succinimidyl ester (CFSE)‐labelled human haemopoietic cells through six cell cycles based on the precise halving of their CFSE‐fluorescence at each mitosis. Using this approach in combination with DNA or surface antigen staining, we show that the addition of Flt3‐ligand (FL) to a cytokine cocktail consisting of Steel factor, IL‐3, IL‐6 and G‐CSF increased the proportion of CD34+ (CD45RA/CD71)−, but not CD34+(CD45RA/CD71)+, human marrow cells initially recruited into division in vitro, shortened the overall cycle time of their progeny, and enhanced the production of a derivative CD34+CD38− population through several (up to four) cell generations. These studies also showed that during the first 4 d there was no detectable apoptosis among the progeny of the CD34+(CD45RA/CD71)− cells generated in the presence of this four‐cytokine cocktail, regardless of the presence of FL. The availability of a technique for monitoring changes in the properties of individual cells as a function of their mitotic history and under conditions where they are asynchronously recruited to divide provides a new and powerful approach for studies of the regulation of primitive human haemopoietic cell proliferation and differentiation.
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一种新型单细胞增殖测定表明，随着时间的推移，长期培养起始细胞 (LTC-IC) 的维持是由一小部分 LTC-IC 的广泛增殖造成的。
影响因子：
20.3
作者：
Verfaillie,CM;Miller,JS
通讯作者：
Miller,JS
影响因子：
15.9
作者：
MUTA, K;KRANTZ, SB;WICKREMA, A
通讯作者：
WICKREMA, A
无血清骨髓培养物中造血功能的维持涉及静态祖细胞的连续募集。
影响因子：
2.6
作者：
Lansdorp,PM;Dragowska,W
通讯作者：
Dragowska,W
DOI：
10.1073/pnas.91.25.12223
发表时间：
1994-12-06
影响因子：
11.1
作者：
SAUVAGEAU, G;LANSDORP, PM;HUMPHRIES, RK
通讯作者：
HUMPHRIES, RK