Application of adenosine in preparation of drugs or reagents for promoting embryo formation or improving embryo quality
An embryo quality and adenosine technology, applied in the field of medicine, can solve the problem of little research on granulosa cell metabolism, and achieve the effect of improving oocyte quality and embryo development, reducing meiotic defects, and decreasing ROS levels.
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Embodiment 1
[0037] Example 1 Effect of Adenosine on Mouse Embryo Development
[0038] Take 8-week-old ICR mice, inject PMSG, inject hCG 48 hours later, and collect materials 16 hours later; obtain hyaluronidase-digested granulosa cells from the oviduct ampulla under a microscope, clean the naked eggs, and distribute them evenly at 37°C for 5 %CO 2 Well-balanced M2 (Aged: control group); M2 added with DMSO (Aged+D (Aged+DMSO): solvent control group) and M2 added with 10 μM adenosine dissolved in DMSO (Aged+A (Aged +Adenosine): In the experimental group) in the culture medium, in vitro fertilization was carried out after 12 hours. Simultaneously, fresh oocytes (Fresh: fresh control group) were taken out immediately for in vitro fertilization. The sperm of the 4 groups for in vitro fertilization all came from the same male mouse. Observing the embryonic development process, and performing fluorescent staining statistics on the blastocysts, it was found that the formation of early embryos ...
Embodiment 2
[0039] Example 2 Changes of intracellular adenosine content and transport carriers after exogenous addition of adenosine
[0040] Take 8-week-old ICR mice, inject PMSG, inject hCG 48 hours later, and collect materials 16 hours later; obtain hyaluronidase-digested granulosa cells from the oviduct ampulla under a microscope, clean the naked eggs, and distribute them evenly at 37°C for 5 %CO 2 Well-balanced M2 supplemented with DMSO (Aged+D (Aged+DMSO): solvent control group), M2 supplemented with 10 μM adenosine dissolved in DMSO (Aged+A (Aged+Adenosine): experimental group 1) and Added M2 (Aged+A+C (Aged+Adenosine+Caffeine): experimental group 2) culture solution with 10 μM adenosine and caffeine dissolved in DMSO, washed the naked eggs with PBS after 12 hours, and collected them. At the same time, oocytes in the Fresh group (Fresh: fresh control group) were immediately taken out, digested with hyaluronidase, washed with PBS, and then collected. The 4 groups of samples were a...
Embodiment 3
[0041] Example 3 Detection of adenosine content in human granulosa cells and follicular fluid and the effect of adenosine on oocyte quality
[0042] The granulosa cells and follicular fluid of the IVF population with normal epidemiological information were collected, grouped according to the embryo quality on the third day, and divided into a high-quality group and a low-quality group with a high-quality embryo rate of 40%. Adenosine content, showing that the low-quality group adenosine content increased in granulosa cells and decreased in follicular fluid ( image 3 ).
[0043] Since the adenosine content in the follicular fluid of the low-quality group decreased, and considering that the follicular fluid is the external environment of the oocyte, we added adenosine (the appropriate concentration was 10 μM) to the in vitro culture medium of the oocyte in the animal experiment. Take 8-week-old ICR mice, inject PMSG, inject hCG 48 hours later, and collect materials 16 hours la...
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