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Rhesus monkeys produced by nuclear transfer

L Meng1, J J Ely, R L Stouffer

  • 1Division of Reproductive Sciences, Oregon Regional Primate Research Center, Beaverton 97006-3499, USA.

Biology of Reproduction
|August 1, 1997
PubMed
Summary

This study demonstrates that nuclear transfer can successfully produce genetically identical rhesus monkeys. Researchers used donor cells from embryos to create reconstituted embryos, which were then transferred to surrogate mothers, resulting in the birth of two healthy infants. This technique provides a valuable new model for medical research.

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Area of Science:

  • Reproductive biology and nuclear transfer technologies
  • Nonhuman primate genetics and developmental biology

Background:

The generation of genetically uniform nonhuman primate models remains a significant challenge for biomedical research. No prior work had successfully demonstrated the viability of cloning rhesus macaques through nuclear transfer techniques. Such models are necessary for investigating heritable physiological parameters in controlled settings. Prior research has shown that cloning is possible in other mammalian species. However, the specific requirements for primate oocyte activation and embryo development differ significantly from other models. That uncertainty drove the need for specialized protocols tailored to macaque biology. This gap motivated the development of the procedures described in this report. Researchers aimed to establish a reliable method for producing identical primates for future gene therapy studies.

Purpose Of The Study:

The aim of this study is to demonstrate that nuclear transfer is a viable technology for producing identical rhesus macaques. Researchers sought to address the lack of genetically uniform primate models for biomedical experimentation. This problem limits the ability to study heritable physiological parameters in a controlled manner. The team hypothesized that using diploid blastomeres as nuclear donors would facilitate successful cloning. They aimed to refine the procedures for oocyte enucleation and activation in this species. By overcoming technical hurdles, the investigators intended to provide a new resource for gene therapy research. The motivation stems from the need for consistent animal subjects in complex genetic studies. This work seeks to validate a methodology that could significantly advance the field of primate developmental biology.

Keywords:
cloningembryo reconstitutionprimate modelsgenetics

Frequently Asked Questions

The researchers utilized nuclear transfer, where diploid blastomeres from embryos were injected into enucleated oocytes. These reconstituted embryos were then fused using electric pulses and cultured on buffalo rat liver cells before being transferred to surrogate mothers to achieve pregnancy.

The team employed cycloheximide to trigger the activation of the oocytes after they were enucleated by removing the first polar body and underlying ooplasm. This chemical agent is essential for initiating the developmental processes required for successful embryo reconstitution in this specific primate model.

The researchers performed enucleation by aspirating the first polar body and the underlying ooplasm to ensure the oocytes were chromosome-free. This technical step is necessary to prevent the inclusion of maternal genetic material, thereby ensuring the resulting embryos carry only the donor nucleus.

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Main Methods:

The review approach involved recovering oocytes from females treated with gonadotropins for maturation. Investigators performed enucleation by removing the first polar body and associated ooplasm from each cell. They activated these cells using exposure to cycloheximide to prepare them for donor nuclei. Individual diploid blastomeres were harvested from embryos produced via in vitro fertilization. These donor cells were placed under the zona pellucida of the prepared oocytes. Fusion was achieved through the application of controlled electric pulses to the reconstituted structures. The resulting embryos underwent coculture on buffalo rat liver cells to support development. Finally, the team performed cryostorage before transferring the embryos into synchronized host mothers.

Main Results:

Key findings from the literature indicate that nuclear transfer successfully produced two live rhesus macaque births. Out of nine females receiving twenty-nine reconstituted embryos, three pregnancies were confirmed by the researchers. The two resulting infants consisted of one male and one female. Genotype analysis at seven highly variable short tandem repeat loci confirmed the identity of the offspring. The donor blastomeres were sourced from embryos that were either fresh or frozen-thawed. This result confirms that both storage methods are compatible with the cloning process. The successful birth rate demonstrates the viability of the described technical pipeline. These findings provide empirical evidence that identical primates can be generated using this specific methodology.

Conclusions:

The authors demonstrate that nuclear transfer serves as a functional technology for generating identical rhesus macaques. Synthesis and implications suggest that these primates offer a robust model for exploring heritable physiological traits. The successful birth of two infants confirms the efficacy of the described embryo reconstitution process. Genotype analysis at seven highly variable loci provides definitive proof of the genetic identity of the offspring. These findings indicate that both fresh and frozen-thawed blastomeres can act as effective nuclear donors. The study highlights the potential for using these models in diverse research activities. Future applications may include advanced gene therapy investigations requiring genetically consistent subjects. This work establishes a foundation for expanding primate cloning research in a controlled manner.

Genotype analysis at seven highly variable short tandem repeat loci served as the data type to confirm parentage. This molecular approach provided unequivocal evidence that the infants were genetically identical to the donor blastomeres rather than the surrogate mothers.

The study measured success through pregnancy rates and live births, achieving two live births from twenty-nine transferred embryos. This measurement demonstrates the feasibility of the technique, contrasting with the failure rates observed in previous attempts to clone primates using different methodologies.

The authors propose that these genetically identical primates provide a powerful model for gene therapy. They suggest that such models are particularly useful for research activities where the physiological parameters under study are heritable, offering a more controlled environment than genetically diverse populations.