Pluripotency and self-renewal of canine embryonic stem cells

犬胚胎干细胞的多能性和自我更新

• 批准号: 250191-2012
• 负责人: Betts, Dean
• 金额: $ 1.89万
• 依托单位: University of Western Ontario
• 依托单位国家: 加拿大
• 项目类别: Discovery Grants Program - Individual
• 财政年份: 2012
• 资助国家: 加拿大
• 起止时间: 2012-01-01 至 2013-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=505703
• 关键词: Pluripotency; self; renewal; canine; embryonic

Stem cells are characterized by their capacity for prolong cell division and their ability to differentiate into multiple cell types. As such, stem cells help preserve tissue functions throughout life. Aerobic metabolism, while enabling efficient energy production, also generates reactive oxygen species (ROS). Stem cell biology has primarily focused on the harmful effects of ROS, which include cell/tissue aging, cancer development and activation of anti-stress repair mechanisms. In contrast, exciting new data that we have contributed to generating, indicates that the characteristic traits of stem cells are in fact regulated by ROS. My NSERC funded research program (continuously since 2001) has resulted in the novel development of several canine embryonic stem cell (cESC) lines. Interestingly, our cESCs appear to require both leukemia inhibitory factor (LIF) and basic fibroblast growth factor (bFGF) in their culture medium to survive, proliferate and maintain their stem cell characteristics - unlike mouse or human ESCs, which solely require LIF or bFGF, respectively. The next critical step in characterizing these novel canine stem cells is to characterize and understand how ROS govern their stem cell attributes. Using canine embryonic stem cell culture models that mimic physiological low and high oxidative conditions, we will characterize the redox status and signaling pathways that control canine embryonic stem cell division in the undifferentiated state. Using a novel protein separation technique, we will determine the spectrum of disulfide-linked proteins that are affected by increased ROS levels and by modulating the levels of ROS aim to increase the efficiency of producing induced pluripotent stem cells from canine fibroblasts. Identification and analysis of ROS sensing proteins will provide insight into how oxidative cellular conditions influence stem cell functions. Support for these studies will place Canada in the forefront of these emerging biotechnologies, providing high-tech training for numerous highly qualified personnel (HQP) while enhancing the biomedical and commercial potential for implementing stem cell therapies in the dog.

干细胞的特点是具有延长细胞分裂的能力以及分化成多种细胞类型的能力。因此，干细胞有助于在整个生命过程中保持组织功能。有氧代谢在实现高效能量产生的同时，也会产生活性氧（ROS）。干细胞生物学主要关注ROS的有害影响，包括细胞/组织衰老、癌症发展和抗应激修复机制的激活。相比之下，我们贡献的令人兴奋的新数据表明干细胞的特征实际上是由 ROS 调节的。 我的 NSERC 资助的研究项目（自 2001 年以来持续进行）已导致几种犬胚胎干细胞 (cESC) 系的新开发。 有趣的是，我们的 cESC 似乎需要在其培养基中同时含有白血病抑制因子 (LIF) 和碱性成纤维细胞生长因子 (bFGF) 才能生存、增殖并维持其干细胞特性 - 与小鼠或人类 ESC 不同，后者分别仅需要 LIF 或 bFGF。表征这些新型犬干细胞的下一个关键步骤是表征和了解 ROS 如何控制其干细胞属性。使用模拟生理低和高氧化条件的犬胚胎干细胞培养模型，我们将表征控制犬胚胎干细胞在未分化状态下分裂的氧化还原状态和信号通路。使用一种新型蛋白质分离技术，我们将确定受 ROS 水平增加影响的二硫键连接蛋白质的谱，并通过调节 ROS 水平来提高从犬成纤维细胞产生诱导多能干细胞的效率。 ROS 传感蛋白的鉴定和分析将有助于深入了解氧化细胞条件如何影响干细胞功能。对这些研究的支持将使加拿大处于这些新兴生物技术的前沿，为众多高素质人员（HQP）提供高科技培训，同时增强在狗身上实施干细胞疗法的生物医学和商业潜力。