METHODS FOR PRODUCTION OF ICSI AND SCNT DERIVED MACAQUE STEM CELLS

ICSI 和 SCNT 衍生的猕猴干细胞的生产方法

• 批准号: 8172703
• 负责人: GERALD SCHATTEN
• 金额: $ 1.92万
• 依托单位: TEXAS BIOMEDICAL RESEARCH INSTITUTE
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-05-01 至 2011-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8172703
• 关键词: Animal Model; Autistic Disorder; Computer Retrieval of Information on Scientific Projects Database; Data; Derivation procedure; Diabetes Mellitus; Disease; Ejaculation; Embryo; Embryo Cloning; Fertilization; Freezing; Funding; Future; Germ Cells; Goals; Grant; Heart Diseases; Immune; Institution; Intracytoplasmic Sperm Injections; Laboratories; Laparotomy; Macaca; Methods; Modeling; Normalcy; Oocytes; Parkinson Disease; Pharmaceutical Preparations; Pilot Projects; Production; Protocols documentation; Research; Research Personnel; Resources; Safety; Source; Staging; Stem cells; Testing; Therapeutic; United States National Institutes of Health; Work; blastocyst; embryo culture; embryonic stem cell; established cell line; human embryonic stem cell; nonhuman primate; pluripotency; rectal; somatic cell nuclear transfer; sperm cell

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. For this protocol, macaques will be hormonally stimulated with super-ovulatory drugs to produce oocytes that will be collected via laparotomy. These oocytes will be fertilized in the laboratory using fresh or frozen sperm collected by rectal probe electro-ejaculation. The collected gametes will be used for fertilization in the laboratory by intracytoplasmic sperm injection and the embryos cultured to the blastocyst stage. Once reaching this stage, several methods will be employed to attempt to derive embryonic stem cells. In addition, some oocytes may undergo somatic cell nuclear transfer to form cloned embryos. These embryos will also be cultured to the blastocyst stage in order to derive stem cells. Long term, we anticipate that this research will enable us to optimize methods for the derivation of macaque embryonic stem cells from both fertilized and SCNT embryos. However, for this pilot project, our goal is to simply obtain data on cyno follicular stimulation and determine whether or not cynos may be an ideal animal model for future work. Any established cell lines will undergo rigorous testing to determine their pluripotency and normalcy. While human embryonic stem cells may have incredible therapeutic value for treating a wide range of disorders such as Parkinson's diabetes, autism, heart disease, Alzheimer's etc., rigorous testing in animal models is necessary before any such trials can begin. Macaques provide a valuable non-human primate model in which to determine the safety and efficacy of immune-matched stem cells.

该子项目是利用该技术的众多研究子项目之一 资源由 NIH/NCRR 资助的中心拨款提供。子项目及 研究者 (PI) 可能已从 NIH 的另一个来源获得主要资金， 因此可以在其他 CRISP 条目中表示。列出的机构是 对于中心来说，它不一定是研究者的机构。 在该方案中，猕猴将受到超排卵药物的激素刺激，产生卵母细胞，并通过剖腹手术收集卵母细胞。 这些卵母细胞将在实验室中使用通过直肠探针电射精收集的新鲜或冷冻精子进行受精。 收集到的配子将用于在实验室通过胞浆内单精子注射进行受精，并将胚胎培养至囊胚阶段。 一旦达到这个阶段，将采用多种方法来尝试获得胚胎干细胞。 此外，一些卵母细胞可能会经历体细胞核移植形成克隆胚胎。这些胚胎还将被培养至囊胚阶段，以获得干细胞。 从长远来看，我们预计这项研究将使我们能够优化从受精胚胎和 SCNT 胚胎中衍生猕猴胚胎干细胞的方法。然而，对于这个试点项目，我们的目标是简单地获得有关食蟹猴卵泡刺激的数据，并确定食蟹猴是否可能是未来工作的理想动物模型。 任何已建立的细胞系都将经过严格的测试，以确定其多能性和正常性。虽然人类胚胎干细胞对于治疗帕金森氏症、糖尿病、自闭症、心脏病、阿尔茨海默氏症等多种疾病可能具有令人难以置信的治疗价值，但在开始任何此类试验之前，必须在动物模型中进行严格的测试。 猕猴提供了一个有价值的非人类灵长类动物模型，可以用来确定免疫匹配干细胞的安全性和有效性。