Efficient Embryo Transfer in the Common Marmoset Monkey (Callithrix jacchus) with a Reduced Transfer Volume: A Non-Surgical Approach with Cryopreserved Late-Stage Embryos1

Abstract

Among primates, the common marmoset is suitable for primate embryology research. Its small body size, however, has delayed the technical development of efficient embryo transfer. Furthermore, three factors have been determined to adversely affect the performance of marmoset embryo transfer: nonsurgical approaches, the use of cryopreserved embryos, and the use of late-stage embryos. Here we performed embryo transfer under conditions that included the above three factors and using either a small (1 μl or less) or a large volume (2-3 μl) of medium. The pregnancy and birth rates were 50% (5/10) and 27% (3/11), respectively, when using the large volume, and 80% (8/10) and 75% (9/12), respectively, when using the small volume. The latter scores exceed those of previous reports using comparable conditions. Thus, it appears that these three previously considered factors could be overcome, and we propose that reducing the transfer volume to 1 μl or less is essential for successful marmoset embryo transfer.

Generation of transgenic marmosets expressing genetically encoded calcium indicators

“…We were able to achieve an embryo cleavage rate of 80.9% and a transgene expression rate of 89.6% after subzonal injection of the lentiviruses, indicating the efficient integration of these proviruses into the marmoset genome with minimal interference on embryonic viability. In addition, recent progresses made on the adaptation of assisted reproductive techniques for marmoset monkeys, including controlled ovarian stimulation protocols, in vitro maturation and fertilization of oocytes, non-invasive sperm collection, embryo collection from donor females and transfer into recipient females, have strongly facilitated the production of transgenic marmosets and enhanced their value as experimental animal models 23 44 45 46 47 48 49 . By implementing these techniques, we were able to repeatedly recover pre-implantation stage marmoset NAT embryos noninvasively without any residual impact to the donor female’s reproductive health.…”

Transgenic Monkey Model of the Polyglutamine Diseases Recapitulating Progressive Neurological Symptoms

“…The animals were subjected to embryo collection on d 4–7 and embryo transfer on d 2–5. Embryo collection and transfer were performed by nonsurgical methods as previously described (Ishibashi et al 2013a, b). Briefly, for embryo collection, a blunt tapered 28-G inner stainless steel cannula and a blunt tapered 19-G outer stainless steel cannula were inserted into the uterus with the aid of ultrasonography.…”

Assisted Reproductive Techniques on South American Wild Mammals

“…In marmoset monkey, due to its small body size, the technical development of an efficient embryo transfer is being delayed. Different factors are being studied, such as nonsurgical approaches, embryo cryopreservation, use of late-stage embryos, and volume medium [112] to overcome these gaps. Using embryos in different developmental stages (10-to 16-cell, morula, and blastocysts) after vitrification in Cryotop, Ishibashi et al [112] defended that reducing the transfer volume to 1 µL or less is essential for successful embryo transfer this species.…”

“…Different factors are being studied, such as nonsurgical approaches, embryo cryopreservation, use of late-stage embryos, and volume medium [112] to overcome these gaps. Using embryos in different developmental stages (10-to 16-cell, morula, and blastocysts) after vitrification in Cryotop, Ishibashi et al [112] defended that reducing the transfer volume to 1 µL or less is essential for successful embryo transfer this species. This procedure provides pregnancy and birth rates of 80% (8/10) and 75% (9/12), while the use of larger volumes (2-3 µL) results in pregnancy and birth rates of 50% (5/10) and 27% (3/11), respectively.…”