scholarly journals Efficient marmoset genome engineering by autologous embryo transfer and CRISPR/Cas9 technology

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Yukiko Abe ◽  
Harumi Nakao ◽  
Motoki Goto ◽  
Moe Tamano ◽  
Michinori Koebis ◽  
...  
Keyword(s):  
Embryo Transfer ◽  
Genome Engineering ◽  
Assisted Reproductive Technologies ◽  
Small Body ◽  
Common Marmoset ◽  
Reproductive Technologies ◽  
Reproductive Capacity ◽  
Primate Species ◽  
Novel Method

AbstractGenetic engineering of non-human primates, which are most closely related to humans, has been expected to generate ideal animal models for human genetic diseases. The common marmoset (Callithrix jacchus) is a non-human primate species adequate for the production of genetically modified animals because of their small body size and high reproductive capacity. Autologous embryo transfer (AET) is routinely utilized in assisted reproductive technologies for humans but not for experimental animals. This study has developed a novel method for efficiently producing mutant marmosets using AET and CRISPR/Cas9 systems. The embryos were recovered from oviducts of naturally mated females, injected with Cas9/guide RNA, and transferred into the oviducts of the donors. This AET method can reduce the time for in vitro culture of embryos to less than 30 min. This method uses an embryo donor as the recipient, thus reducing the number of animals and allowing for “Reduction” in the 3R principles of humane experimental technique. Furthermore, this method can utilize nulliparous females as well as parous females. We applied our novel method and generated the 6 marmosets carrying mutations in the fragile X mental retardation 1 (FMR1) gene using only 18 females including 14 nulliparous females.

scholarly journals Epigenetic Effects of Assisted Reproductive Technology in Human Offspring

2019 ◽  
Author(s):  
Wei Chen ◽  
Yong Peng ◽  
Xinyi Ma ◽  
Siming Kong ◽  
Shuangyan Tang ◽  
...  
Keyword(s):  
Embryo Transfer ◽  
Assisted Reproductive Technologies ◽  
Adult Life ◽  
Reproductive Technologies ◽  
Fresh Embryo Transfer ◽  
Vitro Fertilization ◽  
Nuclear Families ◽  
Genome Wide ◽  
Parental Background

AbstractThe births of more than 8 million infants have been enabled globally through assisted reproductive technologies (ARTs), including conventional in vitro fertilization (IVF) and intracytoplasmic sperm injection (ICSI) with either fresh embryo transfer (ET) or frozen embryo transfer (FET). However, the potential for elevated risks of ART-related disorders persists in adult life, and the underlying epigenetic mechanisms are largely uncharacterized. Here, we recruited 100 nuclear families and profiled the DNA methylomes, genome-wide histone modifications and transcriptomes to clarify the inherent extra risks attributable to specific ART procedures. We discovered that IVF-ET seemed to introduce less disturbance into the infant epigenome than IVF-FET or ICSI-ET did. Furthermore, we noted approximately half of the DNA methylomic changes in ART-conceived offspring could be explained by parental background biases. Through removal of the parental effect, we confirmed that ART per se would introduce minor DNA methylation changes locally. More importantly, we found that ART-induced epigenomic alterations were highly enriched in the processes which might contribute to increased incidence of preeclampsia during pregnancy and metabolic syndrome in offspring. Overall, our study provides an epigenetic basis for the potential long-term health risks in ART-conceived offspring that reinforces the need to review all methods of human ART.

Possibility of elimination of pathogen from Babesia infected animals by means of embryo transfer

2014 ◽  
Vol 11 (2) ◽  
pp. 36-42
Author(s):  
P Erdenetogtokh ◽  
S Ganbat ◽  
Hiroshi Suzuki
Keyword(s):  
Embryo Transfer ◽  
Assisted Reproductive Technologies ◽  
Morphological Characteristics ◽  
Reproductive Technologies ◽  
Development Stage ◽  
Babesia Microti ◽  
Control Group ◽  
Mice Model ◽  
Economic Sectors

Babesia infections occur mainly in animals, and are transmitted by ticks. The severity of the diseases varies considerably depending on the species of Babesia involved as well as the immune response of the infected animal. In Mongolia infection produced by Babesia parasites is widely spread, provoking severe damage to the agricultural and economic sectors. Currently, strategies to control and prevent the infection are inefficient. Indeed, the necessity to look for suitable and accessible strategies to obtain animals free from the infection is needed. Currently, assisted reproductive technologies (ART) are used for the improvement of productivity in livestock. Moreover, embryo transfer seams to be useful approach to obtain clean embryos obtained from infected animals. Therefore, by using a mice model (ICR) infected with Babesia microti, an alternative method to obtain animals free from infection was examined. ICR mice at 8 weeks old were challenged with 0.2 ml of 1x107 IRBC/ml by i.p injection. After infection, superovulation was induced and then embryos were obtained and washed. Then, their development stage along with their morphological characteristics were monitored. In vitro embryos obtained from uninfected mice were used as a control group. The results indicate that the infection does not have any influence on pre-implantation embryonic development and morphological characteristics. Thus, we suggest that embryos obtained from infected animals might be useful for embryo transfer in order to improve productivity of livestock and reduce the risk of congenital infection. In summary, ART such as embryo transfer might be an useful technique in countries where Babesiosis is an endemic disease. DOI: http://dx.doi.org/10.5564/mjas.v11i2.214 Mongolian Journal of Agricultural Sciences Vol.11(2) 2013 pp.36-42

scholarly journals Long-Term Effects Following Fresh/Vitrified Embryo Transfer Are Transmitted by Paternal Germline in a Large Size Rabbit Cohort

Animals ◽  
2020 ◽  
Vol 10 (8) ◽  
pp. 1272 ◽  
Author(s):  
Ximo Garcia-Dominguez ◽  
José Salvador Vicente ◽  
María P. Viudes-de-Castro ◽  
Francisco Marco-Jiménez
Keyword(s):  
Embryo Transfer ◽  
Assisted Reproductive Technologies ◽  
Later Life ◽  
Reproductive Technologies ◽  
Early Embryo ◽  
Embryo Cryopreservation ◽  
Long Term Effects ◽  
Adult Body Weight ◽  
Female Germline

The concept of developmental programming suggests that the early life environment influences offspring phenotype in later life, whose effects may also be manifested in further generations. Valuable pieces of evidence come from the fields applying assisted reproductive technologies (ARTs), which deprive embryos of their optimal maternal environment and were thus associated with subsequent developmental deviations. Recently, we demonstrated that the in vitro manipulations during a vitrified embryo transfer procedure incurs a cumulative and transgenerational decline in the growth performance of the resulting offspring. Here, we provide a longitudinal study to investigate whether previous developmental deviations could be indistinctly paternally or maternally transmitted using crossbred mattings. Our findings revealed that early embryo manipulations through fresh and vitrified embryo transfer incurred paternally transmissible effects over the growth pattern and adult body weight, which seemed not inheritable via the female germline. Similar inheritable effects were observed after fresh and vitrified embryo transfer, suggesting that disturbing optimal embryo development through in vitro manipulations was the principal trigger of transmissible effects, rather than embryo cryopreservation per se.

87 Assessment of pregnancy success following transfer of embryos produced in vitro using frozen - thawed semen from cloned and noncloned Bos indicus bulls

2019 ◽  
Vol 31 (1) ◽  
pp. 169
Author(s):  
O. Sebastián ◽  
F. Guerrero ◽  
R. Romero ◽  
F. Muñoz ◽  
A. Parlange ◽  
...  
Keyword(s):  
Embryo Transfer ◽  
Assisted Reproductive Technologies ◽  
Numerical Range ◽  
Bos Indicus ◽  
Reproductive Technologies ◽  
The State ◽  
In Vitro Production ◽  
Exact Test ◽  
Significant Difference

Assisted reproductive technologies (ART) continue to develop rapidly, allowing for the development of techniques to increase reproductive efficiency and contribute to the genetic improvement of cattle. Some of these techniques include in vitro production (IVP) of embryos and embryo transfer. These modern ART can help produce offspring with highly desirable characteristics. However, there is a lack of information on the percentage of pregnancies obtained following transfer (P/ET) of IVP embryos derived using semen of cloned Bos indicus bulls. The objective of this study was to compare embryo transfer results of IVP embryos created using frozen-thawed semen from 5 Brahman bulls (Bos indicus) with characteristics and genetics of high commercial value. The embryos were produced on two different dates, 45 days apart, using pooled oocytes harvested by ovum pickup from 15 Brahman cows at random stages of the oestrous cycle. Procedures for IVP were performed in a commercial laboratory (Genemex Internacional) in the state of Chiapas, Mexico. For IVF, conventional semen was used from 1 bull (B1) and his clone (B12), the grandson of B1 (B2), and from 2 nonrelated bulls (B3 and B4). A total of 100 embryos were transferred nonsurgically by a private practitioner on a ranch in the state of Campeche, Mexico. The recipients were commercial crossbred cows synchronized using a FTET program. On Day 0, recipients received an intravaginal device containing 1.9g of progesterone (CIDR) and 2mg of oestradiol benzoate IM. On Day 8, the CIDR was removed and cows received 25mg of dinoprost tromethamine, 200IU of eCG, and 0.5mg of oestradiol cypionate IM. Embryos were transferred on Day 17. The overall P/ET was 42% (42/100). The P/ET for IVP embryos produced with semen from bulls B1, B12, B2, B3, and B4 was 3/15 (20%), 3/8 (37%), 23/42 (55%), 8/20 (40%), and 5/15 (33%), respectively. The P/ET was numerically greater for embryos produced using semen from the cloned bull (37%; B12) compared to embryos produced using semen from the original noncloned bull (20%; B1), although this difference was not statistically significant (P=0.62, Fisher’s exact test). There was a significant difference (P<0.05) for the P/ET obtained with embryos produced using semen from bulls B1 and B2, but results for the other bulls were not significantly different. As far as we know, this is the first scientific report in Mexico concerning the use and comparison of semen from cloned and noncloned bulls for the production and transfer of bovine IVP embryos. In general, a wide numerical range of P/ET using the different bulls was observed (i.e. 20-55%). In this preliminary study, there was no impact of using frozen-thawed semen from a cloned bull for IVP on P/ET. The results from this research can contribute to the study and development of ART to improve P/ET obtained using Zebu IVP embryos. However, further research with a larger numbers of animals is required to confirm whether using semen from cloned and noncloned Bos indicus bulls for IVP impacts pregnancy success following embryo transfer.

253. Calcium ionophore induction of marmoset oocyte activation

2005 ◽  
Vol 17 (9) ◽  
pp. 101
Author(s):  
C. G. Grupen ◽  
R. B. Gilchrist ◽  
S. J. Schulz ◽  
L. J. Ritter ◽  
D. T. Armstrong
Keyword(s):  
Assisted Reproductive Technologies ◽  
Cumulus Cell ◽  
Calcium Ionophore ◽  
Reproductive Technologies ◽  
Primate Species ◽  
Oocyte Activation ◽  
Cell Stage ◽  
Developmental Potential ◽  
Parthenogenetic Activation

The marmoset monkey (Callithrix jacchus) is a valuable model for developing assisted reproductive technologies in humans and endangered primate species. Calcium ionophore treatments have been used to induce parthenogenetic activation in a number of species, but the effectiveness of this reagent in initiating marmoset embryo development has not yet been reported. The aim of this study was to determine the developmental potential of in vitro matured (IVM) marmoset oocytes, following treatment with calcium ionophore. Immature oocytes from large (LA; >1.5 mm) and small (SA; 0.67–1.5 mm) antral follicles were isolated from the ovaries of FSH-primed animals and cultured in modified G2 medium for 26–30 h at 37.0°C in 6% CO2 in air. Meiotically mature oocytes were sequentially incubated with 5 μM ionomycin for 5 min and 2 mM 6-dimethylaminopurine for 3 h and cultured in G1/G2 sequential medium at 37.0°C in 5% O2, 6% CO2, 89% N2 for 10 days. Cumulus cell expansion associated with LA oocytes (n=118) was greater than that of SA oocytes (n=212), as determined using well established classification criteria (2.7±0.1 v. 1.8±0.2; P<0.01). A greater proportion of LA oocytes completed meiosis to the metaphase-II stage compared with SA oocytes (85±7% v. 63±7%; P<0.05). Pronuclear formation was induced at similar rates in mature oocytes of both groups, but the rate of cleavage was higher for LA oocytes compared with SA oocytes (93±6% v. 66±5%; P<0.05). The number of cells per embryo was not different between the groups.This is the first study to demonstrate that calcium ionophore effectively induces parthenogenetic activation in IVM marmoset oocytes. However, the development of parthenotes was limited beyond the 8-cell stage. Further studies are needed to determine the cause of the developmental block.

scholarly journals Perinatal outcomes of human singletons conceived naturally versus assisted reproductive technologies: analysis of the effect of stimulated IVF, modified natural IVF, and frozen embryo transfer

2021 ◽  
Vol 26 (1) ◽  
Author(s):  
Adhwaa Khudhari ◽  
Ali Mourad ◽  
Simon Phillips ◽  
Mohammad Zubair Alam ◽  
Robert Hemmings ◽  
...  
Keyword(s):  
Birth Weight ◽  
Embryo Transfer ◽  
Gestational Age ◽  
Assisted Reproductive Technologies ◽  
Perinatal Outcomes ◽  
Reproductive Technologies ◽  
Primary Objective ◽  
Frozen Embryo Transfer ◽  
Significant Difference

Abstract Background Obstetrical outcomes in assisted reproduction techniques (ART) were compared with naturally conceived pregnancies and among each other in multiple reports. However, many important changes in the practice of in vitro fertilization (IVF) over the years, including single embryo transfers (sET) and the introduction of modified natural IVF (mnIVF), and the advances in the frozen embryo transfer (FET) might have impacted the outcomes. Our study is the first to our knowledge to assess four different groups, including spontaneous pregnancies, mnIVF, stimulated IVF (sIVF), and FET altogether in a head-to-head comparison. This is a retrospective study on perinatal outcomes of singleton babies conceived naturally or using three different ART protocols between 2011 and 2014. The primary objective was the comparison of gestational age and birth weight between spontaneously conceived pregnancies (NAT, n= 15,770), mnIVF (n=235), sIVF (n=389), and FET (n=222). Results Our results show a significant difference in favor of naturally conceived pregnancies over ART in term of gestational age. In fact, the gestational age of babies in the NAT group was statistically higher compared to each one of the ART groups alone. Regarding the birth weight, the mean was significantly higher in the FET group compared to the other categories. Conclusion Differences in perinatal outcomes are still found among babies born after different modes of conception. However, there is still need for well-designed high-quality trials assessing perinatal outcomes between naturally conceived pregnancies and different ART protocols based on different maternal and treatment characteristics.

scholarly journals Towards Improving the Outcomes of Assisted Reproductive Technologies of Cattle and Sheep, with Particular Focus on Recipient Management

Animals ◽  
2020 ◽  
Vol 10 (2) ◽  
pp. 293 ◽  
Author(s):  
Jamee Daly ◽  
Hayley Smith ◽  
Hayley A. McGrice ◽  
Karen L. Kind ◽  
William H.E.J. van Wettere
Keyword(s):  
Embryo Transfer ◽  
Assisted Reproductive Technologies ◽  
Effective Means ◽  
Cost Effective ◽  
Reproductive Technologies ◽  
Flock Size ◽  
Phenotypic Marker ◽  
Vitro Fertilization ◽  
Sheep Meat

The Australian agricultural industry contributes AUD 47 billion to the Australian economy, and Australia is the world’s largest exporter of sheep meat and the third largest for beef. Within Australia, sheep meat consumption continues to rise, with beef consumption being amongst the highest in the world; therefore, efficient strategies to increase herd/flock size are integral to the success of these industries. Reproductive management is crucial to increasing the efficiency of Australian breeding programs. The use of assisted reproductive technologies (ARTs) has the potential to increase efficiency significantly. The implementation of multiple ovulation and embryo transfer (MOET) and juvenile in vitro fertilization and embryo transfer (JIVET) in combination with genomic selection and natural mating and AI is the most efficient way to increase genetic gain, and thus increase reproductive efficiency within the Australian livestock industries. However, ARTs are costly, and high variation, particularly between embryo transfer recipients in their ability to maintain pregnancy, is a significant constraint to the widespread commercial adoption of ARTs. The use of a phenotypic marker for the selection of recipients, as well as the better management of recipient animals, may be an efficient and cost-effective means to increase the productivity of the Australian livestock industry.

Development of assisted reproductive technologies for Mus spretus†

2020 ◽  
Author(s):  
Ayumi Hasegawa ◽  
Keiji Mochida ◽  
Shogo Matoba ◽  
Kimiko Inoue ◽  
Daiki Hama ◽  
...  
Keyword(s):  
Embryo Transfer ◽  
Chorionic Gonadotropin ◽  
Assisted Reproductive Technologies ◽  
Laboratory Mouse ◽  
Reproductive Technologies ◽  
Efficient Production ◽  
High Survival ◽  
Mus Spretus ◽  
High Genetic Diversity

Abstract The genus Mus consists of many species with high genetic diversity. However, only one species, Mus musculus (the laboratory mouse), is common in biomedical research. The unavailability of assisted reproductive technologies (ARTs) for other Mus species might be a major reason for their limited use in laboratories. Here, we devised ARTs for Mus spretus (the Algerian mouse), a commonly used wild-derived Mus species. We found that in vitro production of M. spretus embryos was difficult because of low efficacies of superovulation with equine chorionic gonadotropin or anti-inhibin serum (AIS) (5–8 oocytes per female) and a low fertilization rate following in vitro fertilization (IVF; 15.2%). The primary cause of this was the hardening of the zona pellucida but not the sperm’s fertilizing ability, as revealed by reciprocal IVF with laboratory mice. The largest number of embryos (16 per female) were obtained when females were injected with AIS followed by human chorionic gonadotropin and estradiol injections 24 h later, and then by natural mating. These in vivo-derived 2-cell embryos could be vitrified/warmed with a high survival rate (94%) using an ethylene glycol-based solution. Importantly, more than 60% of such embryos developed into healthy offspring following interspecific embryo transfer into (C57BL/6 × C3H) F1 female mice. Thus, we have devised practical ARTs for Mus spretus mice, enabling efficient production of embryos and animals, with safe laboratory preservation of their strains. In addition, we have demonstrated that interspecific embryo transfer is possible in murine rodents.

scholarly journals Long-Term Phenotypic and Proteomic Changes Following Vitrified Embryo Transfer in the Rabbit Model

Animals ◽  
2020 ◽  
Vol 10 (6) ◽  
pp. 1043 ◽  
Author(s):  
Ximo Garcia-Dominguez ◽  
Francisco Marco-Jiménez ◽  
David S. Peñaranda ◽  
José Salvador Vicente
Keyword(s):  
Growth Performance ◽  
Embryo Transfer ◽  
Developmental Plasticity ◽  
Assisted Reproductive Technologies ◽  
Rabbit Model ◽  
Reproductive Technologies ◽  
Long Term Effects ◽  
Mammal Species

Nowadays, assisted reproductive technologies (ARTs) are considered valuable contributors to our past, but a future without their use is inconceivable. However, in recent years, several studies have evidenced a potential impact of ART on long-term development in mammal species. To date, the long-term follow-up data are still limited. So far, studies have mainly focused on in vitro fertilization or in vitro culture, with information from gametes/embryos cryopreservation field being practically missing. Herein, we report an approach to determine whether a vitrified embryo transfer procedure would have long-term consequences on the offspring. Using the rabbit as a model, we compared animals derived from vitrified-transferred embryos versus those naturally conceived, studying the growth performance, plus the weight throughout life, and the internal organs/tissues phenotype. The healthy status was assessed over the hematological and biochemical parameters in peripheral blood. Additionally, a comparative proteomic analysis was conducted in the liver tissue to investigate molecular cues related to vitrified embryo transfer in an adult tissue. After vitrified embryo transfer, birth weight was increased, and the growth performance was diminished in a sex-specific manner. In addition, vitrified-transferred animals showed significantly lower body, liver and heart weights in adulthood. Molecular analyses revealed that vitrified embryo transfer triggers reprogramming of the liver proteome. Functional analysis of the differentially expressed proteins showed changes in relation to oxidative phosphorylation and dysregulations in the zinc and lipid metabolism, which has been reported as possible causes of a disturbed growth pattern. Therefore, we conclude that vitrified embryo transfer is not a neutral procedure, and it incurs long-term effects in the offspring both at phenotypic and molecular levels. These results described a striking example of the developmental plasticity exhibited by the mammalian embryo.

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