Maintenance of Stem Cell Pluripotency in Pigs with Interleukin-6

项目摘要 猪在生理、解剖学和免疫学上都与人类相似。因此，他们服务于 作为一个伟大的生物医学研究模型。然而，无法产生胚胎干细胞 (ESCs)和体细胞重编程诱导的多能干细胞(IPSCs)限制了 这种模式物种在研究人类疾病和再生医学方面的作用。 克服这一限制将增加产生转基因品系的机会 可以用来研究人类疾病。这项工作的长期目标是建立 维护猪ESCs和IPSCs的创新方法，无需 结构性地表达与多能性相关的转录因子。创新战略 在这项探索性方案中测试的是使用白细胞介素6(IL6)来维持脑内的多能性 猪细胞。白介素6还没有作为一种多功能因子在猪身上进行测试，但所有 有迹象表明，它将担任这一职务。以前的报告和最近的调查结果提供了 令人信服的证据表明，IL6在猪、人和其他动物中起着多能因子的作用 哺乳动物。这项提案中的工作将检验IL6是可塑性的驱动因素和 猪ESCs和IPSCs的多能性。计划的目标有两个。目标1将探讨 IL6作用于猪ICM细胞。前提是IL6将保持一个自我更新的种群 PICM细胞类似于幼稚ESCs的真正的多能性特征。目标2将 检测是否可以通过补充IL6建立无转基因猪IPSC。 在IL6存在的情况下对猪成纤维细胞进行重新编程有望减少对 促进多能性的核心多能因子和microRNAs的转基因表达 猪。

Project Summary Pigs are physiologically, anatomically and immunologically similar to humans. Thus, they serve as a great biomedical research model. However, the inability to generate embryonic stem cells (ESCs) and somatic cell reprogrammed induced pluripotent stem cells (iPSCs) has limited the usefulness of this model species for studying human diseases and regenerative medicine. Overcoming this limitation will enhance opportunities to generate genetically-engineered lines that can be used for studying human diseases. The long-term goal of this work is to establish innovative approaches for maintaining porcine ESCs and iPSCs without the need to constitutively express pluripotency-related transcriptional factors. The innovative strategy tested in this exploratory proposal is using interleukin-6 (IL6) to maintain pluripotency in porcine cells. Interleukin-6 has not been tested as a pluripotency factor in pigs, but all indications are that it will serve in this capacity. Previous reports and recent findings provide compelling evidence that IL6 serves as a pluripotency factor in pigs, humans and other mammals. The work in this proposal will test the hypothesis that IL6 is a driver of plasticity and pluripotency in porcine ESCs and iPSCs. Two aims are planned. Aim 1 will explore the actions of IL6 acts on porcine ICM cells. The premise is that IL6 will maintain a self-renewing population of pICM cells that resemble the authentic pluripotency features of naïve ESCs. Aim 2 will examine whether transgene-free porcine iPSC can be established by IL6 supplementation. Reprogramming porcine fibroblasts in the presence of IL6 is anticipated to reduce the need for transgene expression of core pluripotency factors and microRNAs that facilitate pluripotency in pigs.