PRODUCTION OF MHC-DEFINED AND GENETICALLY IDENTICAL MODELS

这个子项目是许多研究子项目中的一个 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得了主要资金， 因此可以在其他CRISP条目中表示。所列机构为 研究中心，而研究中心不一定是研究者的研究机构。 目的：1）开发使用卵裂球分离创造遗传相同猴的策略，2）使用辅助生殖技术繁殖MHC定义的猴家族。 非人灵长类动物是研究人类疾病的有价值的模型。 然而，动物之间固有的变异性减少了研究的好处，使用基因相同的动物可以最大限度地减少这种变异性。 我们的主要目标是制造出同卵双胞胎恒河猴。从控制性卵巢刺激后的卵泡中吸出卵母细胞进行体外受精产生胚胎。 双胞胎是通过在4- 14-细胞阶段的卵裂球分离获得的， Ca，-Mg培养液和卵裂球重新聚集成2个空的透明细胞。分裂后成功增殖的胚胎进行腹腔镜移植给接受孕酮植入或注射的受体。 然而，这些分裂的胚胎没有获得足月妊娠。 增加了一项额外的试验，通过钻孔和卵裂球抽吸对卵裂期胚胎进行活检，以确定胚胎操作对妊娠结局的影响。 在22例注射孕酮补充剂的活组织检查胚胎移植中，4例健康的单胎婴儿是由非手术经子宫移植后的活组织检查胚胎产生的。植入前卵裂球活检可能是一个有价值的方法，以确定特定的遗传性状在个别灵长类动物胚胎作为进一步的信息恒河猴基因型与生理性状的联系。 本项目使用了Animal、CPI和Assay Services。 一份出版物待定。 注：与艾滋病有关。

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. Objective: To 1) develop strategies for creating genetically identical monkeys using blastomere separation and 2) propagate a family of MHC-defined monkeys using assisted reproductive technologies. The nonhuman primate is a valuable model for examining human disease. The benefits to research, however, are reduced by the inherent variability among animals, which could be minimized by the use of genetically identical animals. Our primary goal was to produce identical rhesus macaque twins. Embryos were produced by in vitro fertilization of oocytes aspirated from follicles after controlled ovarian stimulation. Twins were derived by blastomere separation at the 4- to 14-cell stage in Ca, -Mg media and the blastomeres were reaggregated into 2 empty zona pellucida. Embryos that successfully proliferated after splitting underwent laparoscopic transfer to recipients receiving progesterone implants or injections. However, no term pregnancies were obtained with these split embryos. An additional trial with cleavage stage embryos biopsied by zona drilling and blastomere aspiration was added to define the effect of embryo manipulation on pregnancy outcome. Of 22 transfers of biopsied embryos with injected progesterone supplementation, four healthy singleton infants were produced from biopsied embryos following nonsurgical, transcervical transfer. Preimplantation blastomere biopsy could be a valuable approach for defining specific genetic traits in individual primate embryos as further information on the linkage of rhesus genotype with physiological traits is obtained. This project used Animal, CPI, and Assay Services. A publication is pending. Note: AIDS-related.