Quantitative morphology of induced phenotypes in cultured cells and tissues

我们继续与Minoru Ko博士（LG-NIA）合作，利用转录因子操纵来表征小鼠胚胎干细胞诱导的发育途径。我们能够建立一个分化试验基于形态学单独，使用相衬成像没有特定的标记。我们能够证明，随着时间的推移，分化可以被追踪，分化细胞之间的形态相似性在早期就建立起来，并在分化的几天内保持下来。一个重要的里程碑是，我们能够建立一个条件，细胞系显示一致的基因功能分组在几个独立的实验。此外，我们能够扩展这一分析，以比较56个细胞系的形态。这些结果正在准备发表。

We have continued our work in collaboration with Dr. Minoru Ko (LG-NIA) to characterize developmental pathways induced in mouse embryonic stem cells using transcription factor manipulation. We were able to establish a differentiation assay based on morphology alone, using phase-contrast imaging without specific markers. We were able to show that differentiation can be tracked over time, and that morphological similarities between differentiating cells are established early and maintained for several days of differentiation. An important milestone is that we were able to establish a conditions where cell lines showed consistent grouping by gene function over several independent experiments. Additionally, we were able to expand this analysis to compare the morphologies of 56 cell lines. These results are being prepared for publication. We have continued work characterizing the molecular basis of morphological age-state transitions during the C. elegans life-span. In previous published work we used WND-CHARM to identify distinct morphological aging states in C. elegans. We used this technique to sort worms based on their age state during a transition period where an aging population is evenly divided between individuals in Stage I and Stage II. The worms were identical genetically, by chronological age, by growth conditions, and by visual appearance, and could only be sorted into age-states using WND-CHARM. Micro-array experiments performed on these two sub-populations revealed several hundred genes with significantly altered expression profiles. By comparing our gene lists with those from other aging studies in C. elegans, we were able to identify several gene families and functional groups that were unique to our study, as well as several important aging genes that were identified previously. The results of this study are being prepared for publication. In the next stage of this study, we will begin to screen through our gene list using RNAi libraries to shift the timing of the Stage I - Stage II transition.