Abstract
The assisted reproductive technologies (ARTs) have been used in the production of rhesus monkey offspring at the Oregon National Primate Research Center (ONPRC) and that experience is summarized here. Additionally these technologies serve as a source of oocytes/embryos for monozygotic twinning, embryonic stem (ES) cell derivation and cloning. High fertilization efficiencies were realized with conventional insemination or following the use of intracytoplasmic sperm injection (ICSI) and approximately 50% of the resulting embryos grew in vitro to blastocysts. Both fresh and frozen sperm were employed in fertilization by ICSI and the resulting embryos could be low temperature stored for subsequent thawing and transfer when a synchronized recipient female was available or after shipment to another facility. Following the transfer of up to 3 embryos, an overall pregnancy rate of 30% was achieved with increasing rates dependent upon the number of embryos transferred. Singleton pregnancy outcomes following the transfer of ART produced embryos were similar to those observed in a control group of animals in the timed mated breeding colony at ONPRC. ICSI produced embryos were used in efforts to create monozygotic twins by blastomere separation or blastocyst splitting. While pregnancies were achieved following the transfer of demi-embryos, only one was a twin and it was lost to spontaneous abortion. ICSI produced embryos have also served as the source of blastocysts for the derivation of embryonic stem cells. These pluripotent cells hold potential for cell based therapies and we consider the monkey an important translational model in which to evaluate safety, efficacy and feasibility of regenerative medicine approaches based on the transplantation of stem cell-derived progeny. Finally, efforts to produce genetically-identical monkeys by nuclear transfer have been briefly summarized.
Highlights
Notable progress has been realized in the past 20 years in both the development and application of the assisted reproductive technologies (ARTs) in monkeys with advances in ovarian follicular stimulation, fertilization by intractyoplasmic sperm injection (ICSI), culture medium development for the production of blastocysts, low temperature storage of sperm and embryos and embryo manipulation including twinning, cloning and the derivation of embryonic stem cells
This effort has achieved the first twin pregnancies in rhesus monkeys, the first non-human primate infants produced by nuclear transfer of embryonic cells, the first rhesus monkey infant born following the transfer of an ICSI-produced blastocyst employing a non-surgical procedure, the first monkey live birth resulting from the transfer of a demi-embryo created by blastomere separation at the 2-cell stage or blastocyst bisection and the first infants produced following laparoscopic embryo transfer
Based on the results summarized in this mini-review, the ARTs in non-human primates would seem to be ready for widespread application in the propagation of selected, desirable genotypes
Summary
In the past 15 years, research into and practice of the ARTs in rhesus monkeys at ONPRC has resulted in a number of notable accomplishments. Based on the results summarized in this mini-review, the ARTs in non-human primates would seem to be ready for widespread application in the propagation of selected, desirable genotypes This is not to say that there is no longer room for protocol development. Modest improvements in outcome to 50% pregnancy rates following the transfer of 2 embryos can bring that cost down to $4,000 This is based on the expectation that a single controlled ovarian stimulation, oocyte collection, embryo production cycle can support 7–8 embryo transfers and 3–4 offspring. The ARTs may soon be used extensively in the routine propagation of valuable animals These considerations assume that the technical support and the resources already are in place and are based on modest subsidies for ARTCore services at ONPRC. The logistics of providing such services to the wider NHP community could involve the use of one or more centralized facilities or a larger number of local units
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