On the symmetry of siblings: automated single-cell tracking to quantify the behavior of hematopoietic stem cells in a biomimetic setup
On the symmetry of siblings: automated single-cell tracking to quantify the behavior of hematopoietic stem cells in a biomimetic setup
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DOI：
10.1016/j.exphem.2011.10.009
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发表时间：
2012-02-01
影响因子：
2.6
通讯作者：
Roeder, Ingo
中科院分区：
文献类型：
--
作者：
Scherf, Nico;Franke, Katja;Roeder, Ingo
造血干/祖细胞(HSPC)与其局部微环境之间的相互作用是造血组织的关键机制。为了定量研究这一过程，在单细胞水平上对细胞动力学进行时间分辨分析是必不可少的前提。产生足够数量的适当数据的一种方法是使用延时视频显微镜的自动单细胞跟踪。我们描述并应用了新开发的计算算法，允许以高时间和高空间分辨率自动生成单细胞特征的高含量数据，以及重建和统计评估完整的系谱历史。这种方法已经应用于生物工程培养条件下纯化的原代人HSPC的具体例子。系谱信息和细胞特性的动态特征相结合，确定了兄弟姐妹HSPC在细胞周期时间、迁移速度和生长动力学方面的显著对称性。此外，我们证明了HSPC兄弟姐妹的这种对称性可以通过局部仿生微环境的外源线索来改变。以HSPC在仿生培养系统中的生长为例，我们表明，我们的方法为在确定的体外条件下定量分析动态单细胞特征提供了一个有价值的工具，允许整合功能和系谱数据。我们方法的效率和准确性为对发展模式的组织原则和外部线索的各自影响铺平了道路，这些组织原则并不局限于对初级HSPC的研究。(C)2012年ISEH--血液和干细胞学会。由爱思唯尔公司出版。
The interplay between hematopoietic stem and progenitor cells (HSPC) and their local micro-environment is a key mechanism for the organization of hematopoiesis. To quantitatively study this process, a time-resolved analysis of cellular dynamics at the single-cell level is an essential prerequisite. One way to generate sufficient amounts of appropriate data is automatic single-cell tracking using time-lapse video microscopy. We describe and apply newly developed computational algorithms that allow for an automated generation of high-content data of single-cell characteristics at high temporal and spatial resolution, together with the reconstruction and statistical evaluation of complete genealogical histories. This methodology has been applied to the particular example of purified primary human HSPCs in bioengineered culture conditions. The combination of genealogical information and dynamic profiles of cellular properties identified a marked symmetry between sibling HSPCs regarding cell cycle time, but also migration speed and growth kinetics. Furthermore, we demonstrate that this symmetry of HSPC siblings can be altered by exogenous cues of the local biomimetic microenvironment. Using the example of HSPC growth in biomimetic culture systems, we show that our approach provides a valuable tool for the quantitative analysis of dynamic single-cell features under defined in vitro conditions, allowing for integration of functional and genealogical data. The efficiency and accuracy of our approach pave the way for new and intriguing insights into the organizational principles of developmental patterns and the respective influence of exogenous cues not limited to the study of primary HSPCs. (C) 2012 ISEH - Society for Hematology and Stem Cells. Published by Elsevier Inc.