Characterising embryonic stem cell-derived tenocytes and defining the changing role of scleraxis during tendon development

主题：健康生物科学为了帮助成人肌腱再生和减少再损伤的频率，正在开发新的策略，利用干细胞有可能分化成肌腱细胞。然而，在正常的成人肌腱修复过程中，已经有大量的肌腱细胞涌入产生疤痕组织。因此，重要的是设计细胞疗法，可以模仿胎儿的小细胞和再现无疤痕再生。我们已经证明，马的ESCs在注射到受伤的马肌腱后分化为腱细胞。使用体外3D培养系统生成人工肌腱，我们已经确定转化生长因子β 3和机械力是分化的关键驱动因素。然而，esc衍生的小细胞是代表成人细胞还是胎儿细胞尚不清楚。在这个博士学位中，学生将通过对esc衍生的、成人和胎儿的小细胞进行全球基因表达分析来探索新的工作方式。生物信息学方法将用于确定胚胎干细胞所代表的发育阶段。全球基因表达分析也将在敲低硬化轴水平的细胞上进行，以确定硬化轴调控下游的基因。我们的合成细胞培养系统将用于生成人造肌腱，学生将有机会通过使用生物工程技术对肌腱施加力来改进该模型，以更好地模拟体内情况。该项目的结果将为未来的实验和临床研究提供新的信息，以应用细胞为基础的疗法来调节肌腱再生。ENWW:-数据驱动生物学（生物信息学）-全球基因表达分析-细胞生物成像-合成生物学（3D培养，细胞基因工程）-各种实验数据的统计分析

Theme: Bioscience for HealthTo aid adult tendon regeneration and reduce the frequency of re-injury novel strategies are being developed which use stem cells that have the potential to differentiate into tenocytes. However, during normal adult tendon repair there is already a substantial influx of tenocytes which produce scar tissue. It is therefore important to design cell therapies that can mimic fetal tenocytes and recapitulate scar-less regeneration. We have shown that equine ESCs differentiate into tenocytes following their injection into the injured horse tendon. Using an in vitro 3D culture system to generate artificial tendons, we have identified transforming growth factor beta 3 and mechanical force as key drivers of differentiation. However, whether ESC-derived tenocytes represent adult or fetal cells is unknown. In this PhD the student will exploit new ways of working by performing global gene expression analyses on ESC-derived, adult and fetal tenocytes. Bioinformatics approaches will be used to determine the developmental stage that ESC-tenocytes represent. Global gene expression analyses will also be performed on cells that are engineered to have knocked down scleraxis levels to identify genes that are downstream of scleraxis regulation. Our synthetic cell culture system to generate artificial tendons will be used and the student will have the opportunity to refine this model by employing bioengineering techniques to apply forces to the tendons to better mimic the in vivo situation. The results of this project will provide novel information to inform future experimental and clinical studies on the application of cell based-therapies for modulating tendon regeneration. ENWW:- data driven biology (bioinformatics)- global gene expression analyses- biological imaging of cells - synthetic biology (3D culture, cell genetic engineering)- statistical analysis of various experimental data