Abstract
As an antioxidant, procyanidin B1(PB1) can improve the development of somatic cell nuclear transfer (SCNT) embryos; PB1 reduces the level of oxidative stress (OS) during the in vitro development of SCNT embryos by decreasing the level of reactive oxygen species (ROS) and increasing the level of glutathione (GSH) and mitochondrial membrane potential (MMP). Metabolite hydrogen peroxide (H2O2) produces OS. Catalase (CAT) can degrade hydrogen peroxide so that it produces less toxic water (H2O) and oxygen (O2) in order to reduce the harm caused by H2O2. Therefore, we tested the CAT level in the in vitro development of SCNT embryos; it was found that PB1 can increase the expression of CAT, indicating that PB1 can offset the harm caused by oxidative stress by increasing the level of CAT. Moreover, if H2O2 accumulates excessively, it produces radical-(HO-) through Fe2+/3+ and damage to DNA. The damage caused to the DNA is mainly repaired by the protein encoded by the DNA damage repair gene. Therefore, we tested the expression of the DNA damage repair gene, OGG1. It was found that PB1 can increase the expression of OGG1 and increase the expression of protein. Through the above test, we proved that PB1 can improve the repairability of DNA damage. DNA damage can lead to cell apoptosis; therefore, we also tested the level of apoptosis of blastocysts, and we found that PB1 reduced the level of apoptosis. In summary, our results show that PB1 reduces the accumulation of H2O2 by decreasing the level of OS during the in vitro development of SCNT embryos and improves the repairability of DNA damage to reduce cell apoptosis. Our results have important significance for the improvement of the development of SCNT embryos in vitro and provide important reference significance for diseases that can be treated using SCNT technology.
Highlights
Somatic cell nuclear transfer (SCNT) is a technology that transfers the nuclei of nonpluripotent differentiated somatic cells into mature oocytes, so that the transplanted oocytes have pluripotency and can develop into normal individuals
In regard to the total blastocyst cell numbers in SCNT embryos of cultured with KSOM medium supplemented with 0 and 50 μM PB1, the result showed that the grou3pof 13 with supplemented 50 μM PB1 total blastocyst cell numbers were significantly increased compared with the control group (93.86 ± 17.52 vs. 76.00 ± 10.18, p < 0.01, Figure 1E,F)
To determine the DNA damage repairability of PB1, we detected the OGG1 mRNA and protein expression in blastocysts; the results showed that the OGG1 mRNA expression was significantly increased and protein expression was increased in the 50 μM PB1 group compared with the control group (114.27 ± 11.86 vs. 79.12 ± 24.82 pixels per embryo, p < 0.05, Figure 3D,E, respectively)
Summary
Somatic cell nuclear transfer (SCNT) is a technology that transfers the nuclei of nonpluripotent differentiated somatic cells into mature oocytes, so that the transplanted oocytes have pluripotency and can develop into normal individuals. Scientists used their experience in rhesus monkeys to obtain fibroblasts using fetuses or infants [3] Both normal humans and type 1 diabetes (T1D) patients have produced ntESCs (nuclei of ntESC cell lines as donors and nuclei of adult somatic cells as donors for nuclear transfer) [4,5]. Reproductive cloning has great potential to expand the population of important agricultural economic animals and save endangered animals without sacrificing donor animals [6,7,8,9]. Another potential application of SCNT technology is the generation of new animal models for human diseases [10,11]. Because of the huge application potential of nuclear transfer, in addition to improving the efficiency of nuclear transfer from the perspective of epigenetic modification, it is very important to explore other aspects to improve the efficiency of nuclear transfer
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
