Introduction of HEPES medium commonly used in in vitro fertilization (IVF) CAS NO.1185-53-1

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In vitro fertilization (IVF), a successful technology in the 1950s, has developed rapidly in the last 20 years and has become an important and conventional animal reproduction biotechnology. HEPES, Chinese name hydroxy4-ethylpiperazine ethanethiosulfonic acid, as an important medium, is mainly used to prevent the rapid change of medium pH. Under the open culture condition, when observing the cells, the culture medium is separated from the environment of 5% CO2, CO2 gas escapes rapidly and the pH rises rapidly. If HEPES is added, the pH can be maintained at about 7.0. Generally, HEPES should be added during clonal culture.

In animal IVR technology, healthy sperm with strong vitality will be screened from semen for further in vitro fertilization (IVF) and intrauterine insemination (IUI). When in use, sperm cleaning solution (such as bicarbonate or HEPES buffer) is added to the sperm injection chamber and the screened sperm chamber to fill the channel, and then the sperm stock solution is added to the sperm injection chamber and incubated at 37 ℃ for 30 minutes. Healthy sperm with strong vitality can enter the screened sperm chamber through the micropores on the microfluidic filter membrane.

During in vitro fertilization, mature oocytes and sperm need to be taken out from the body, cultured in vitro for a period of time, and then artificial insemination. In these processes, due to the separation from the mother's conditions, various culture solutions need to be used to simulate the physiological environment in the body. Today, let's introduce some common culture media used in the process of in vitro fertilization:

Oocyte culture:

At present, the research on bovine oocyte maturation in vitro mainly focuses on taking TCM199 as the basic culture medium, and then adding hormones, serum, follicular fluid, growth factors and granulosa cells to simulate the environment of oocyte maturation in vivo and the interaction between various components, so as to achieve the maturation effect equivalent to that in vivo. Common formula of oocyte maturation medium: the basic medium is TCM-199, supplemented with 10% FCS, 25mm HEPES, 0.55mg/ml sodium pyruvate and 100iu penicillin and streptomycin;

Recommended formula of sperm cell culture medium:

Sperm washing solution: Bo / TALP basic solution + 6mg / ml BSA (fraction V) + 5mm caffeine sperm capacitation solution: Bo / TALP basic solution + 6mg / ml BSA (fraction V) + 5mm caffeine + 60ug / ml heparin.

Note: HEPES and other organic buffers can effectively adjust the pH of the medium and are suitable for a variety of cell lines. In fact, some standard medium formulations contain HEPES. However, HEPES may be toxic, especially to some differentiated cells, so its effect should be evaluated before use.

In vitro fertilization is of great significance to the study of animal reproductive mechanism, animal husbandry, medicine and the protection of endangered animals.. In the improvement of livestock varieties, in vitro fertilization technology provides a cheap and efficient means for embryo production, and HEPES, as an indispensable material in the culture medium, is of great value in making full use of excellent variety resources, shortening the breeding cycle of livestock and accelerating the speed of variety improvement.