Development of pluripotent cell lines from Livestock, and animal models for pharmacology/toxicology applications

通过基于iPSC（诱导多能干细胞）的技术将细胞重编程为多能胚胎状态，彻底改变了细胞疗法和药物开发的发展前景。将这些进展成功转化为动物研究模型和牲畜，在开发基于细胞的实验平台方面具有重要的应用，用于研究健康生物体的遗传和分子基础，生产性状和抗病性，以及疾病进展，干预策略，药物测试和毒理学。不幸的是，到目前为止，非鼠或人细胞的重编程策略取得了有限的成功。然而，在理解重编程的要求和促进扩展多能干细胞扩增的培养系统的开发方面的最新进展意味着产生和利用重编程的动物细胞现在是可能的1 -3。该项目的总体目标是使用这些当代最先进的方法开发重编程动物细胞，验证其分化潜力并研究其效用。最初，该项目将专注于桑格研究所的刘彭涛教授提供的新型iPSCs。这些细胞是使用这些新技术制造的，并已用于该项目。评估扩展多能干细胞的自我更新特性，并将该技术转化为从其他物种（包括用于药理学/毒理学应用的动物）获得iPSC。这将检验以下假设：这些新技术对于重新编程和促进来自不同物种的干细胞的繁殖更有效，并且有助于理解这些细胞内的细胞信号传导机制的调节。评估这些新的多能细胞的分化潜力，并确定是否可以通过遗传修饰进行操作。这将检验功能丧失和获得实验可以指导来自这些新细胞系的特定祖细胞的编程和扩增的假设。这将涉及使用CRISPR基因编辑来生成谱系标记细胞（例如肌肉、肠道、巨噬细胞）以监测分化。该项目将补充基于细胞的系统的应用，以询问来自动物育种实验的丰富遗传数据，作为罗斯林研究所新BBSRC研究所战略计划的一部分，并将扩大Censo生物技术公司iPSC相关专业知识的范围。

Reprogramming of cells to a pluripotent embryonic state through iPSC (induced pluripotent stem cell) based technologies has revolutionised prospects for the development of cellular therapies and drug development. Successful translation of these advances to animal research models and livestock has important applications in developing cell-based experimental platforms for studying the genetic and molecular basis of the healthy organism, production traits and disease resistance -as well as disease progression, intervention strategies, drug testing and toxicology. Unfortunately, until now reprogramming strategies for non-murine or human cells has met with limited success. However, very recent advances in understanding the requirements for reprogramming and the development of culture systems that promote the expansion of extended pluripotent stem cells have meant that generating and exploiting reprogrammed animal cells is now possible1-3. The overall aim of this project is to develop reprogrammed animal cells using these contemporary state-of-the-art methodologies, characterise their differentiation potential and investigate their utility. Initially the project will focus on novel iPSCs provided by Professor Pentao Liu at the Sanger Institute. These cells have been created using these novel technologies and have been made available for the project.The objectives of the project are:1. Evaluate the self-renewal properties of the extended pluripotent stem cells and translate this technology to the derivation of iPSCs from other species - including animals used in pharmacology/toxicology applications. This will test the hypothesis that these novel technologies are more effective for reprogramming and promoting propagation of stem cells from different species and contribute to understanding the regulation of cellular signalling mechanisms within these cells.2. Evaluate the differentiation potential of these novel pluripotent cells and determine if this can be manipulated through genetically modification. This will test the hypothesis that loss and gain of function experiments can direct the programming and expansion of specific progenitor cells from these novel cell lines. This will involve the use of CRISPR gene editing to generate lineage marked cells (e.g. muscle, gut, macrophage) to monitor differentiation.This project will complement the application of cell based systems for interrogating the wealth of genetic data from animal breeding experiments, as part of the Roslin Institutes new BBSRC Institute Strategic Programme, and will broaden the scope of the iPSC-related expertise at Censo biotechnologies.