Research articleMinimally invasive transabdominal collection of preimplantation embryos from the common marmoset monkey (Callithrix jacchus)
Introduction
The common marmoset (Callithrix jacchus), a small New World primate originating from northeastern Brazil, is being increasingly used as a nonhuman primate (NHP) model in the field of biomedical sciences, including neuroscience, stem cell research, drug toxicology, immunology, and reproductive biology. Use of the marmoset has several advantages, including its small body size, prolificacy, and strong genetic similarity to humans [1].
Adult marmosets range in height from 25 to 35 cm and weigh approximately 350 to 500 g. The animal's small size offers advantages, such as low feeding costs, easy handling and retention, and relatively small breeding space requirements. Importantly, marmosets have several features facilitating the study of reproductive and developmental biology, such as early sexual maturation (12 to 18 mo), a relatively short gestation period (approximately 144 days), and postpartum ovulation and conception. Furthermore, the female marmoset has high fecundity; females usually give birth to twins and may produce 40 to 80 offspring during their lifetime. Additionally, the marmoset's ovarian cycle can be synchronized using prostaglandins, facilitating time- and cost-efficient embryo collection in a group of animals [2], [3]. Collectively, these advantages render the marmoset a highly suitable species for production of genetically modified nonhuman primates for human disease models. In that regard, transgenic marmosets carrying the green fluorescent protein gene with germ line transmission have been produced from naturally mated fertilized embryos and in vitro-fertilized embryos [4]. Numerous marmoset embryos are required to propagate this transgenic marmoset production technique and clarify marmoset embryonic development.
An efficient abdominal incision method for embryo collection in marmosets has been developed, but this method is invasive and animals required at least 2 mo for recovery [5]. A nonsurgical method has also has been described, but was difficult to reproduce [6]. Therefore, a new method for minimally invasive embryo collection is desirable. The overall objective of this study was to develop a new minimally invasive technique (transabdominal method) for collecting naturally conceived marmoset preimplantation embryos and to compare it with a more invasive technique (standard abdominal incision method) for collecting naturally conceived embryos from common marmosets. To assess the reproducibility of the newly established method, its applicability and success rate (i.e., embryo recovery) were tested at two research centers.
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Animals
Approval for the study procedures was obtained from the Institutional Animal Care and Use Committee, and the study was performed in accordance with the guidelines of the Central Institute for Experimental Animals (CIEA). Experiments in Germany were conducted under reference number AZ 33.42502–066/06.
Adult common marmosets (>2 yr old) were purchased from CLEA Japan (Tokyo, Japan), whereas the German marmosets were obtained from a self-sustaining breeding colony of approximately 500 animals at
Comparison of the abdominal incision and transabdominal methods
At CIEA, embryo recovery rates were 48.0% for abdominal incision method and 48.4% for the transabdominal method (Table 1). At DPZ, recovery rates were 88.9% for abdominal incision method and 77.8% for the transabdominal method (Table 1). Overall, at least one ovulation product was recovered in 50.3% of abdominal incision method and 49.4% of the transabdominal method attempts. The abdominal incision method yielded an average of 1.56 ± 0.06 (mean ± SD) ovulation products per flush at CIEA and
Discussion
We developed a new embryo collection method and compared the results with those of the abdominal incision method reported by Summers et al. [5]. The disadvantages of abdominal incision procedures include scar formation, inflammation and, in some animals, adhesions of the ovary and uterus to the peritoneum. Previous reports have described nonsurgical techniques for recovery of uterine-stage preimplantation embryos from the common marmoset monkey [6], though this method was difficult for us to
Acknowledgments
This study was supported by the Strategic Research Program for Brain Sciences; the Global COE Program for Education and Research Centre for Metabolomic Systems Biology; the Funding Program for World-Leading Innovative R&D on Science and Technology (FIRST) from the Ministry of Education, Culture, Sports, Science and Technology (MEXT); and Precursory Research for Embryonic Science and Technology (PRESTO) funding from the Japan Science and Technology Agency (JST). It was also supported by the
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Contributed equally to this work.