PRODUCTION OF MHC-DEFINED AND GENETICALLY IDENTICAL MODELS

这个子项目是许多研究子项目中利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 目的：1)开发利用卵裂球分离创造基因完全相同的猴子的策略；2)利用辅助生殖技术繁殖由MHC定义的猴子家族。 非人灵长类动物是研究人类疾病的有价值的模型。然而，研究的好处由于动物之间固有的变异性而减少，这种变异性可能会因使用基因相同的动物而最小化。我们的主要目标是培育出同卵的恒河猴双胞胎。体外受精取自卵泡的卵母细胞，经控制性卵巢刺激后获得胚胎。在钙镁培养液中，在4-14-细胞期通过卵裂球分离获得双胞胎，并将卵裂球重新聚集成2个空的透明带。分裂后成功增殖的胚胎接受了腹腔镜下的移植，接受了孕酮植入或注射。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，用卵裂期胚胎通过透明带钻孔和卵裂球抽吸活组织检查，以确定胚胎处理对妊娠结局的影响。在22个注射孕酮补充的活组织胚胎移植中，有4个健康的单胎婴儿是从非手术、经宫颈移植后的活组织胚胎中产生的。随着对恒河猴基因与生理性状联系的进一步了解，植入前卵裂球活检可能是确定个体灵长类胚胎中特定遗传性状的有价值的方法。 这个项目使用了动物、CPI和化验服务。 一份出版物正在等待发布。 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.