Quantitative Analysis of Melanocyte Migration in vitro Based on Automated Cell Tracking under Phase Contrast Microscopy Considering the Combined Influence of Cell Division and Cell-Matrix Interactions
Quantitative Analysis of Melanocyte Migration in vitro Based on Automated Cell Tracking under Phase Contrast Microscopy Considering the Combined Influence of Cell Division and Cell-Matrix Interactions
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DOI：
10.1051/mmnp/20105101
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发表时间：
2010-01-01
影响因子：
2.2
通讯作者：
Xavier, F.
中科院分区：
文献类型：
--
作者：
Letort, V.;Fouliard, S.;Xavier, F.
本研究的目的是描述和分析黑素细胞在体外迁移的规律和时空动态，以及它与细胞分裂和与基质的相互作用。黑素细胞谱系特别有趣，因为它与胚胎发育和肿瘤发生/转移(黑色素瘤)有关。生物学实验是在从野生型和β-连环素转基因小鼠(BCAT*)建立的两个黑素细胞系上进行的。众所周知，多功能的β-连环蛋白分子能够调节与细胞增殖和迁移有关的各种基因的转录，特别是在黑素细胞谱系中。我们还研究了纤维连接蛋白，一种结合整合素的细胞外基质蛋白，从而为细胞提供粘附点并鼓励迁移。随着单个细胞的迁移被跟踪，需要自动化的方法来处理由延时视频显微镜产生的大量数据。一种基于模型的自动细胞跟踪方法在样本上进行了评估，并与手动跟踪进行了比较。这种方法被认为是研究整体细胞行为的可靠方法。对所有观察到的序列的应用提供了对影响黑素细胞体外迁移的因素的洞察：突变形式的β-连环蛋白的黑素细胞显示出更高的分裂率，并且由此导致的细胞密度的增加不会引起接触性生长抑制。然而，细胞密度限制了细胞位移的幅度，尽管它们的运动性受到的影响较小。与底物结合的高纤维连接蛋白浓度促进了细胞的迁移和运动，对野生型细胞的影响比对β-连环蛋白过度表达的细胞更强烈。在分裂过程中，细胞迁移速度迅速增加，而后缓慢下降。对这些数据的分析预计将导致生物学上的答案，并为未来更好的建模过程提供一个框架。
The aim of this study was to describe and analyze the regulation and spatio-temporal dynamics of melanocyte migration in vitro and its coupling to cell division and interaction with the matrix. The melanocyte lineage is particularly interesting because it is involved in both embryonic development and oncogenesis/metastasis (melanoma). Biological experiments were performed on two melanocyte cell lines established from wild-type and beta-catenin-transgenic mice (bcat*). The multi-functional beta-catenin molecule is known to be able to regulate the transcription of various genes involved in cell proliferation and migration, particularly in the melanocyte lineage. We also investigated fibronectin, an extra-cellular matrix protein that binds integrins, thereby providing adhesion points for cells and encouraging migration. As the migration of individual cells were followed, automated methods were required for processing the large amount of data generated by the time-lapse video-microscopy. A model-based approach for automated cell tracking was evaluated on a sample by comparison with manual tracking. This method was found reliable in studying overall cell behaviour. Its application to all the observed sequences provided insight into the factors affecting melanocyte migration in vitro: melanocytes of mutated form of beta-catenin showed higher division rates and no contact inhibition of growth was induced by the resulting increase in cell density. However, cell density limited the amplitude of cell displacements, although their motility was less affected. The high fibronectin concentration bound to substratum promoted cell migration and motility, the effect being more intense for wild-type cells than for cells with beta-catenin over-expression. During the division process, cell migration speed increased rapidly then decreased slowly. Analyses of such data is expected to lead both to biological answers and to a framework for a better modeling processes in the future.