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: 8357681
• 负责人: GERALD SCHATTEN
• 金额: $ 0.26万
• 依托单位: TEXAS BIOMEDICAL RESEARCH INSTITUTE
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-05-01 至 2012-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8357681
• 关键词: Alzheimer' s Disease; Animal Model; Autistic Disorder; Data; Derivation procedure; Diabetes Mellitus; Disease; Ejaculation; Embryo; Embryo Cloning; Fertilization; Freezing; Funding; Future; Germ Cells; Goals; Grant; Heart Diseases; Immune; Intracytoplasmic Sperm Injections; Laboratories; Laparotomy; Macaca; Methods; Modeling; National Center for Research Resources; Normalcy; Oocytes; Parkinson Disease; Pharmaceutical Preparations; Pilot Projects; Primates; Principal Investigator; Production; Protocols documentation; Research; Research Infrastructure; Resources; Safety; Source; Staging; Stem cells; Testing; Therapeutic; United States National Institutes of Health; Work; blastocyst; cost; 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. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. 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.

这个子项目是许多利用资源的研究子项目之一 由NIH/NCRR资助的中心拨款提供。子项目的主要支持 子项目的主要研究者可能是由其他来源提供的， 包括其他NIH来源。 列出的子项目总成本可能 代表子项目使用的中心基础设施的估计数量， 而不是由NCRR赠款提供给子项目或子项目工作人员的直接资金。 对于该方案，将用超排卵药物对猕猴进行生殖刺激以产生卵母细胞，将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。 收集的配子将用于在实验室中通过卵胞浆内单精子注射进行受精，并将胚胎培养至囊胚期。 一旦达到这个阶段，将采用几种方法来尝试获得胚胎干细胞。 此外，一些卵母细胞可以进行体细胞核移植以形成克隆胚胎。这些胚胎也将被培养到胚泡阶段，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和SCNT胚胎中获得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关于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可能具有令人难以置信的治疗价值，用于治疗广泛的疾病，如帕金森氏症、糖尿病、自闭症、心脏病、阿尔茨海默氏症等，在任何这样的试验开始之前，必须在动物模型中进行严格的测试。 猕猴提供了一种有价值的非人灵长类动物模型，用于确定免疫匹配干细胞的安全性和有效性。