Establishment of novel Expanded Potential Stem Cells (EPSCs) from bovine pre-implantation embryos: in vitro and in vivo studies.

从牛植入前胚胎中建立新型扩展潜能干细胞（EPSC）：体外和体内研究。

• 批准号: 465494419
• 负责人: Dr. Monika Nowak-Imialek
• 金额: --
• 依托单位: Klinikum rechts der Isar der Technischen Universität München (MRI)
• 依托单位国家: 德国
• 项目类别: Research Grants
• 资助国家: 德国
• 项目状态: 未结题
• 来源: https://gepris.dfg.de/gepris/projekt/465494419?language=en
• 关键词: Establishment; novel; Expanded; Potential; Stem

In many species including humans, rats, mice and monkeys embryonic stem cells (ESCs) can be derived from the inner cell mass of pre-implantation embryos, so-called blastocysts. These cell lines can self-renew and grow in vitro for indefinite time. However, despite many efforts, bona fide pluripotent stem cell lines could not yet be derived from cattle. While some progress has been made, reported bovine ESC lines do not meet the stringent criteria for pluripotency, i.e. the capacity to form germ line chimeras. Recently, embryonic stem cell lines were derived from porcine embryos using novel, so-called “expanded culture conditions”. These cells, named Expanded Potential Stem Cells (EPSCs), possess the special characteristics of being able to differentiate not only into different cell types of the body but also into extraembryonic tissue. The aim of the proposed research is to establish bovine Expanded Potential Stem Cells (bEPSCs) from in vitro produced embryos. These cells should remain undifferentiated indefinitely under appropriate in vitro culture conditions and at the same time possess the ability to differentiate into any cell type of the body in vitro and in vivo, including their contribution to the germ line and eventually the production of functional gametes. In the proposed studies, we plan to use a novel Expanded Potential Stem Cell Medium (EPSCM) to establish bovine embryonic stem cell lines. Embryonic stem cells will be isolated from 8-to-16-cell-stage embryos and from blastocysts (Objective 1) and characterized by morphology, in vitro growth, karyotype stability, pluripotency gene expression, accessibility for genetic manipulation and differentiation potential in vitro (Objective 2). We anticipate that by using EPSC culture conditions we will be able to establish bovine Expanded Potential Stem Cell (bEPSC) lines. These stem cells would be similar to pre-implantation blastomers, with the ability to generate germline chimeras after re-introduction into bovine pre-implantation embryos (Objective 3). The proposed work will provide unique bovine stem cell lines with expanded potential that can be great candidates to introduce multiple genetic modifications and enable the generation of transgenic animal models. These cell lines with expanded potential to differentiate into all embryonic and extraembryonic cell types could be of great interest for reproductive biotechnology as well as for innovative in vitro breeding methods.

在包括人类、大鼠、小鼠和猴子在内的许多物种中，胚胎干细胞（ESCs）可以从植入前胚胎的内部细胞团中获得，即所谓的囊胚。这些细胞系可以自我更新，并在体外无限期生长。然而，尽管做了许多努力，真正的多能干细胞系还不能从牛身上获得。虽然已经取得了一些进展，但报道的牛ESC系不符合多能性的严格标准，即形成生殖系嵌合体的能力。最近，利用新型的所谓“扩大培养条件”从猪胚胎中获得了胚胎干细胞系。这些细胞被命名为扩增潜能干细胞（EPSCs），它们不仅能够分化成身体的不同细胞类型，而且还能分化成胚胎外组织。拟议研究的目的是从体外产生的胚胎中建立牛扩增潜能干细胞（bEPSCs）。这些细胞应在适当的体外培养条件下保持不分化，同时具有在体外和体内分化为人体任何细胞类型的能力，包括它们对生殖系的贡献，并最终产生功能配子。在本研究中，我们计划使用一种新型的扩增潜能干细胞培养基（EPSCM）来建立牛胚胎干细胞系。胚胎干细胞将从8- 16细胞阶段的胚胎和囊胚中分离出来（目标1），并具有形态学、体外生长、核型稳定性、多能性基因表达、遗传操作的可及性和体外分化潜力（目标2）。我们预计，通过使用EPSC培养条件，我们将能够建立牛扩增潜能干细胞（bEPSC）系。这些干细胞将类似于植入前胚泡，在重新导入牛植入前胚胎后能够产生种系嵌合体（目的3）。这项工作将提供独特的牛干细胞系，具有扩大的潜力，可以作为引入多种遗传修饰和产生转基因动物模型的伟大候选者。这些细胞系具有扩大分化为所有胚胎和胚胎外细胞类型的潜力，可能对生殖生物技术以及创新的体外育种方法有很大的兴趣。