Method for improving development quality of in vitro fertilization female embryos after implantation

An in vitro fertilization and embryo technology, applied in embryo cells, biochemical equipment and methods, microorganisms, etc., can solve problems such as solutions that do not achieve expected results, reduce the development efficiency of bovine IVF embryos, and reduce the risk of sex ratio imbalance. The effect of reducing the chance of abnormal fetal development and increasing the chance

Active Publication Date: 2016-06-08
CHINA AGRI UNIV
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Problems solved by technology

Adding 6-phosphate dehydrogenase inhibitors DHEA and 6-AN to the bovine IVF embryo culture medium can significantly improve the sex ratio of bovine IVF blastocysts, but this addit

Method used

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  • Method for improving development quality of in vitro fertilization female embryos after implantation
  • Method for improving development quality of in vitro fertilization female embryos after implantation

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] Embodiment 1 mouse in vitro fertilization embryo production

[0021] 1. In vitro fertilization

[0022]ICR female mice aged 7-8 weeks were intraperitoneally injected with 5IUPMSG (Ningbo Second Hormone Factory), and 47-48h later injected with 5IUhCG (Ningbo Second Hormone Factory). Thirteen hours after the injection of hCG, the mice were sacrificed by cervical dislocation, and the oviducts were dissected and cut, placed in the preheated 37°C M2 operating solution (Sigma Company), and the enlarged part of the oviducts was cut with a syringe under a stereomicroscope to collect cumulus oocytes The complex (mature MⅡ stage oocyte) was transferred to M2 operating solution preheated at 37°C and fully washed before use. Adult healthy male rats (10-12 weeks old) were killed by decapitation, the abdominal cavity was cut open with sterilized scissors, the cauda epididymis was taken, and the sperm in the cauda epididymis was squeezed out with forceps and scissors, and put into a ...

Embodiment 2

[0032] Embodiment 2 bovine in vitro fertilization embryo production

[0033] 1. In vitro fertilization

[0034] Mature MⅡ stage oocytes obtained by hormonal superovulation were washed 3 times in fertilization fluid (BO fluid + 10 mM caffeine + 3 mg / ml BSA) and then put into well-balanced 50 μl fertilization fluid droplets (15 pieces / drop ); Sperm capacitation was carried out by the flotation method, the sperm were floated in the semen solution (BO solution + 20 μg / ml heparin sodium + 6 mg / ml BSA) for 20-30 minutes, and then 600-800 μl of the supernatant was taken and put into a 1.5ml centrifuge tube for centrifugation (1500 rpm , centrifuged for 5 min), the supernatant was discarded, and the washing was repeated once. Remove the supernatant after centrifugation, then add the semen washing solution, count the sperm, then take 50 μl of the treated semen and add it to the fertilization fluid that has been placed in the oocyte, and the final concentration of sperm is 1×10 6 Sper...

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Abstract

The invention provides a method for improving development quality of in vitro fertilization (IVF) female embryos after implantation and correcting sex ratio of IFV embryos. The method comprises: transferring mouse fertilized embryos obtained by IVF technology into a culture solution added with low-dose retinoic acid (RA) for short-term culturing, a female specific apparent modification event (namely x-chromosomal inactivation) in early development of IFV embryos is established normally so that an abnormal development ratio of mouse IFV female embryos after implantation is reduced and the mouse IVF birth sex ratio imbalance problem is finally corrected. In addition, the method is successfully applied in a bovine IVF system, effectively increasing development rate of bovine IFV female blastulas and correcting the sex ratio of bovine IFV blastulas. The RA in the invention is used as a fallopian tube to widely express an endogenous active matter, this matter is suitably added in the IVF system with endogenous ingredients not blindly added, and application safety is greatly ensured.

Description

technical field [0001] The invention relates to the technical field of sexual reproduction, in particular to a method for improving the developmental quality of in vitro fertilized female embryos after implantation. Background technique [0002] In vitro fertilization (invitrofertilization, IVF; test-tube baby) is widely used in the treatment of human infertility. At present, more than 5.4 million cases of test-tube babies have been born in the world, and the rate of 350,000 cases per year is increasing rapidly. In addition, IVF is also widely used in the multiplication of fine breeds of livestock. For example, the proportion of IVF embryos in bovine embryo transfer has reached 40%. Although IVF has such a broad application space, there are still many restrictive factors in IVF, such as: low growth rate, pregnancy loss, premature birth, birth defects, imbalanced sex ratio, and increased risk of diabetes and hypertension. At present, for these IVF defects, mainly by changin...

Claims

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Application Information

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IPC IPC(8): C12N5/073
CPCC12N5/0604C12N2500/30
Inventor 田见晖谭琨安磊吴中红郭敏
Owner CHINA AGRI UNIV
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