In vitro fertilization in mammals

Summary

In vitro fertilization (IVF) is a technological process in which fertilization is accomplished in an in vitro environment between a mature oocyte and an energized sperm, either in vivo or in vitro. In biology, an animal obtained after the transfer of an IVF embryo to the mother is called a test-tubeanimal. This technology is not only of theoretical significance for the study of gamete development, maturation, fertilization and early embryonic cell differentiation, but also of great significance for the solution of human infertility and other social problems, and also of wide practical value for the acceleration of animal reproduction in production.

Principle

The principle of in vitro fertilization is based on the incubation of the oocyte with sperm from the epididymis in the uterine fluid, where the release of the dipolar body and the onset of oogenesis can be observed. However, the spermatozoa are only capable of fertilization after they have been in the female reproductive tract for a certain period of time, a phenomenon known as sperm capacitation.

Appliance

In vitro fertilization is commonly used for research on gamete development, maturation, fertilization and early embryonic cell differentiation, solving social problems such as human infertility, and accelerating animal reproduction.

Operation method

In vitro fertilization in mammals

Principle

The principle of in vitro fertilization is based on the incubation of the oocyte with sperm from the epididymis in the uterine fluid, where the release of the dipolar body and the onset of oogenesis can be observed. However, the spermatozoa are only capable of fertilization after they have been in the female reproductive tract for a certain period of time, a phenomenon known as sperm capacitation.

Materials and Instruments

Equipment:
① Disposable sterile gloves
② Incubator
③ Syringe
Reagents:
① Materials: donor and recipient animals
② Mature culture medium
③ Flushing solution: Duchenne's Phosphate Buffer (PBS), Brinster's medium-3, Synthetic Oviductal Fluid (SOF), Whitten's medium, Ham's F-10 and TCM-199.

Move

The basic process of in vitro fertilization can be divided into the following steps:
I. Acquisition of oocytes
(1) Oocytes from luminal follicles (2-8 mm) are aspirated with a syringe.
(2) Alternatively, the ovary is grasped manually from the rectum, and an aspiration needle is inserted through a puncture in the vaginal wall to aspirate oocytes from a large follicle with the aid of B-mode ultrasound image guidance.
(3) After the ovary is removed from the body, it should be promptly transported to the laboratory for egg culture.
Maturation of oocytes in vitro
(1) Oocytes collected from supernumerary ovulation are already mature in vivo and can be fertilized with sperm directly without culture, while immature oocytes need to be cultured and matured in vitro.
(2) When immature oocytes are cultured, the collected oocytes are first selected and washed under a solid microscope, and then cultured in a maturation medium.
(3) After the oocytes were transferred into small drops, they were cultured in a carbon dioxide incubator under the conditions of 39 ℃, 100% humidity and 5% carbon dioxide in air.
Sperm fertilization in vitro
The combination of caffeine and heparin can greatly increase the rate of fertilization of eggs.
IV . In vitro fertilization or sperm microinjection
A suspension of sperm is added after the oocytes have been equally distributed into small drops in a petri dish.

Caveat

(1) After the ovaries are removed from the body, they should be transported to the laboratory for egg culture in a timely manner, and it is generally advocated that it should be done in 1-4 hours, if it takes more than 6-7 hours, it will significantly reduce the rate of in vitro maturation and fertilization. The temperature at which the ovaries are transported and preserved should preferably be controlled at 25-37 ℃, with a minimum of 20 ℃. Otherwise, it will cause oocyte "cold shock", resulting in a decrease in the maturation rate and fertilization rate.(2) Compared with conventional in vitro fertilization, fertilization and developmental rates are lower in the injectable method.

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