Optimization of red blood cells production from iPS cells using CombiCult Technology

我的博士项目将侧重于利用Plasticell公司的Combicult技术改进人类诱导多能干细胞（human induced pluripotent stem cells, hiPSCs）向红细胞分化的过程。hiPS细胞分化为红细胞是一个复杂的过程，很难在体外重现。在这个过程中涉及的主要问题是了解哪些旁分泌和自分泌信号，以及细胞因子和其他生长因子，参与了红细胞的分化。为了在体外实现更好的红细胞分化，我的实验室与Plasticell公司合作，使用他们的一项新技术——Combicult。该技术将允许在细胞分化过程中使用不同的培养基、细胞因子和生长因子，并使用组合筛选来了解将hiPS分化为成熟红细胞的最佳条件。在这个项目的开始，作为精准医学iCase项目的一部分，我将在Plasticell实习三个月，担任研究员。这次实习将让我了解Combicult技术流程是如何工作的，包括将hiPS细胞放入Combicult技术的过程，以及使用公司开发的生物信息学组合方法确定最佳方案。在实习结束时，预计将有必要的方法来促进hiPS细胞分化为红细胞。目前，实验室正在进行的主要项目之一是研究来自hiPS的巨噬细胞的红细胞岛，以及这种结构的形成和细胞间的接触如何支持脐带血细胞产生红细胞。对于该项目的第二部分，了解使用Combicult技术获得的细胞形成红细胞岛是否促进红细胞的分化和/或成熟将是很有趣的。本研究将在工业方法确定的分化条件验证后在实验室进行。所建立的方法的特性也将在福雷斯特实验室可用的不同iPS细胞系中进行测试。我的项目主要是在小法国的Lesley Forrester实验室的苏格兰再生医学中心（SCRM）进行的。在为期三年半的博士课程期间，作为SCRM研究生课程的一部分，我将参加研讨会、讨论小组、科学介绍、讲习班和期刊俱乐部。作为SCRM博士研究生培训的一部分，我将提交一份为期10周的报告，并将我的项目提案和结果作为研究所内部研讨会的一部分，我将提交一份为期10个月的项目报告，并在年底（5月或6月）介绍项目进展情况。

My PhD project will focus on the improvement of the differentiation process of red blood cells from human induced pluripotent stem cells (hiPSCs) using Plasticell's Combicult technology. Differentiation of hiPS cells into erythrocytes is a complex process proven difficult to recapitulate in vitro. The main issues that are involved in this process is the understanding of which paracrine and autocrine signals, along with cytokines and other growth factors, that are involved in the differentiation of red blood cells. In the aim to achieve better differentiation of erythroid cells in vitro, my laboratory has developed a collaboration with Plasticell Company to use one of their new technologies - Combicult. This technology will allow the use of different media, cytokines and growth factors on cell differentiation process and use a combinatorial screen to understand which is the optimal condition to differentiate hiPS in to mature erythroid cells. At the beginning of this project, and part of the Precision Medicine iCase programme, I will have a three months internship at Plasticell as a researcher. This internship will allow me to comprehend how Combicult technology process works, including the procedure with the hiPS cells into the Combicult technology and the identification of the best protocol using bioinformatics combinatorial method developed by the company. At the end of the internship, it is expected to have the necessary methodology to promote hiPS cells differentiation into red blood cells. At the moment, one of the main projects ongoing in the lab is the study of erythroid islands from hiPS derived macrophages, and how the formation of this structure and the cell-cell contact endorse the generation of erythroid cells from cord blood cells. For the second part of the project, would be interesting to understand if the formation of erythroid islands, with cells obtained using Combicult technology, facilitate the differentiation and/or maturation of erythroid cells. This study would be placed in the lab after validation of differentiation conditions that the industrial approach has identified. Characterisation of the established method will be also tested in different iPS cell lines available in the Forrester lab.My project is mostly based in the Scottish Centre for Regenerative Medicine (SCRM), in Little France, in Lesley Forrester Lab. During this 3.5 year of PhD programme and as part of the post-graduate programme in SCRM I will be attending to seminars, discussion groups, science inductions, workshops and journal clubs. As part of the training for PhD student based at the SCRM, I will submit a 10 week report and present my project proposal and results as part of the internal seminars of the institute, I will submit a 10 month report of the project and present the project evolution at the end of the year (May or June).