A NOVEL APPROACH TO GERM-LIINE PRESERVATION IN MACAQUES

猕猴种系保存的新方法

• 批准号: 6759465
• 负责人: STUART A MEYERS
• 金额: $ 13.1万
• 依托单位: UNIVERSITY OF CALIFORNIA AT DAVIS
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-06-15 至 2006-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6759465
• 关键词: Macaca mulatta; age difference; cell bank /registry; cell sorting; cryopreservation; laboratory mouse; preservation; sperm; stem cell transplantation; stem cells; technology /technique development; tissue /cell culture; xenotransplantation

The non-human primate is a highly relevant model of human disease. Their preservation as important biomedical resources is critical. There is currently a need for populations of genetically-defined monkeys for biomedical investigation in the fields of transplant biology, stem cell biology, transgenics, gene therapy, infectious disease and vaccine development. In addition, the non-human primate model would be a highly valuable model for restoration of spermatogenesis in patients undergoing chemotherapy and radiation therapy. Isolation of spermatogonial stem cells before and autologous transplantation after treatment could be an efficient technique to reverse infertility. The long-term objective of the proposed research is to preserve and reconstitute genotypically unique non-human primates. Immediate objectives will be to develop a successful method to isolate and enrich non-human primate germ line precursor cells and to culture, cryopreserve, and transplant spermatogonial stem cells from genetically unique male macaques to immunodeficient mice and recipient male macaques. These objectives will be accomplished by investigation of the isolation and culture requirements of isolated macaque spermatogonia. We will then determine the specific characteristics of spermatogonial proliferation and differentiation of donor cells from rhesus and cynomolgus macaques transplanted to mouse recipients. We hypothesize that macaque sperm can be banked and then harvested from male mice or recipient macaques by semen collection and subsequently used in artificial insemination and IVF programs at the Regional Primate Research Centers in the US. This project represents a collaboration between Dr. Stuart Meyers, University California at Davis, and Dr. Ina Dobrinski, University of Pennsylvania. All experiments involving live macaques and in-vitro experiments involving selection, culture and freezing of macaque cells will be performed at UC Davis in the laboratory of Dr. Meyers. Experiments involving transplantation of macaque sperm cells into mouse testes will be performed at the University of Pennsylvania in the laboratory of Dr. Dobrinski and experiments involving germ cell transplantation into male macaques will be performed at UC Davis. We expect to expand this study into a fully-independent project.

非人类灵长类动物是人类疾病的高度相关模型。它们作为重要的生物医学资源的保存至关重要。目前，需要在移植生物学，干细胞生物学，转基因，基因治疗，传染病和疫苗发育领域的遗传定义猴子种群进行生物医学研究。此外，非人类灵长类动物模型将是恢复接受化学疗法和放射治疗患者的精子发生的高度有价值模型。在治疗后自体移植之前的精子干细胞分离可能是一种有效的技术来逆转不育。拟议的研究的长期目标是保存和重建基因型独特的非人类灵长类动物。直接目标 将开发一种成功的方法，以分离和富集非人类灵长类动物生殖系的前体细胞，以及培养，冷冻水果和移植精子干细胞从遗传独特的雄性猕猴到免疫缺陷的小鼠和受体男性猕猴。这些目标将通过研究孤立猕猴精子的隔离和培养要求来实现。然后，我们将确定从恒河猴和细胞猕猴猕猴移植到小鼠受体中的精子增生和分化的特定特征。我们假设可以通过精液收集从雄性小鼠或接受者猕猴收获猕猴的精子，然后在区域灵长类动物的人造授精和IVF程序中使用 美国的研究中心。该项目代表了斯图尔特·迈耶斯（Stuart Meyers）博士之间的合作 加州大学戴维斯大学和宾夕法尼亚大学的Ina Dobrinski博士。所有涉及现场猕猴的实验和涉及选择，培养和冻结猕猴细胞的体外实验的实验将在Meyers博士实验室的UC Davis进行。将猕猴细胞移植到小鼠睾丸中的实验将在宾夕法尼亚大学Dobrinski博士实验室进行，并将在UC Davis进行涉及生殖细胞移植到男性猕猴中的实验。我们希望将这项研究扩展到一个完全独立的项目中。