NEW APPROACHES TO THE GENERATION OF GENETICALLY VALUABLE RHESUS MONKEY

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所在的机构。 最初尝试使用猪尾猕猴作为恒河猴胚胎的替代受体，结果表明，怀孕的效率太低，无法常规使用这种方法。为了确保这不是由于在该物种中进行胚胎移植的固有问题，我们开发了一种体外受精系统来产生猪尾胚胎，然后将其用于胚胎移植到猪尾雌性中。这导致了两个猪尾猕猴婴儿的出生。 该项目的重点现在已经转移到使用植入前遗传学诊断（PGD）在胚胎移植到受体之前对其进行基因分型。在过去的一年中，我们进行了6次卵母细胞收集，从中获得182个卵母细胞。总共对120个胚胎进行了活组织检查，并对每种情况下的单个细胞进行PCR分析以确定其性别。每批中的一些胚胎被保存在培养物中，以确保它们的发育没有受到影响。我们目前正在使用性别胚胎进行胚胎移植。最初尝试将胚胎移植回获得卵母细胞的同一只雌性体内，但没有成功。其原因尚不清楚，但可能源于在抽吸过程中卵泡细胞的消耗，这是维持妊娠所必需的。

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. Initial attempts to use pigtail macaques as surrogate recipients for rhesus macaque embryos showed that the efficiency with which pregnancies could be generated was too low to use this method routinely. To ensure that this was not due to inherent problems with performing embryo transfer in this species, we developed an in vitro fertilization system to generate pigtail embryos and then used those in embryo transfers into pigtail females. This resulted in the birth of two pigtail macaque infants. The focus of this project has now shifted towards using preimplantation genetic diagnosis (PGD) to genotype embryos prior to their transfer into recipients. During the past year we performed six oocyte collections from which 182 oocytes were obtained. A total of 120 of the resulting embryos were biopsied and in each case a single cell was subjected to PCR analysis to determine its sex. A few embryos from each batch were kept in culture to ensure their development had not been compromised. We currently are performing embryo transfers using sexed embryos. Initial attempts to transfer embryos back into the same female from which the oocytes had been obtained proved unsuccessful. The reasons for this are not clear but may possibly stem from a depletion of follicular cells during the aspiration process which are necessary to maintain pregnancy.