A NOVEL APPROACH TO GERM-LIINE PRESERVATION IN MACAQUES

非人灵长类动物是与人类疾病高度相关的模型。作为重要的生物医学资源，保存它们是至关重要的。目前，在移植生物学、干细胞生物学、转基因、基因治疗、传染病和疫苗开发等领域，需要对遗传定义的猴子群体进行生物医学研究。此外，非人灵长类动物模型将是一个非常有价值的模型，用于恢复接受化疗和放射治疗的患者的精子发生。治疗前分离精原干细胞，治疗后自体移植是逆转不育症的有效方法。这项拟议研究的长期目标是保护和重建基因独特的非人类灵长类。近期目标 将开发一种成功的方法来分离和丰富非人类灵长类生殖系前体细胞，并从遗传独特的雄性猕猴培养、冷冻保存和移植精原干细胞到免疫缺陷小鼠和受体雄性猕猴。这些目标将通过对分离的猕猴精原细胞的分离和培养要求的调查来实现。然后，我们将确定猕猴和食蟹猴供体细胞移植到小鼠受者体内的精原细胞增殖和分化的特异性。我们假设猕猴的精子可以被储存起来，然后通过收集精液从雄性小鼠或受体猕猴身上获得，然后用于地区灵长类动物的人工授精和试管受精项目 美国的研究中心。这个项目代表了斯图尔特·迈耶斯博士和 加州大学戴维斯分校和宾夕法尼亚大学的艾娜·多布林斯基博士。所有涉及活猕猴的实验和涉及猕猴细胞选择、培养和冷冻的体外实验都将在加州大学戴维斯分校迈耶斯博士的实验室进行。涉及猕猴精子细胞移植到小鼠睾丸的实验将在宾夕法尼亚大学多布林斯基博士的实验室进行，涉及生殖细胞移植到雄性猕猴的实验将在加州大学戴维斯分校进行。我们希望将这项研究扩展为一个完全独立的项目。

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.