Why is there no glucose in the nutrients in the culture medium of animal embryos

Glucose is an energy substance necessary for in vitro culture of somatic cells, but it has adverse effects on the in vitro development of early embryos in hamsters [1], mice [2], cattle [3], sheep [4], and humans [5]. Studies have found that when most animal embryos develop to the mulberry embryo stage, glucose begins to be utilized, and if not added in time, it will inhibit the formation of blastocysts. It can be seen that glucose plays an inhibitory role in the early stage of embryonic development and a promoting role in the later stage.

However, the mechanism by which glucose inhibits early embryonic development is not well understood. There are two main views in the academic community: (1) a crabtree-like effect occurs [4,10].

According to this view, the presence of glucose and phosphate activation enhance glycolysis, which inhibits mitochondrial oxidative metabolism, and the lack of energy ultimately leads to developmental arrest. This view is somewhat representative, but the relationship between glucose phosphate and 2-cell arrest and its fundamental mechanism have not been elucidated. and (2) glucose-induced oxidative stress [3,11-12].

This idea that excess glucose can cause oxidative stress in cells and cause damage to embryonic development has been supported by some studies, but it is still controversial, and there have been no reports of direct measurement of reactive oxygen species (ROS) at all stages of embryonic development in glucose culture. **: Journal of the Second Military Medical University, No. 08, 2009.

Glucose cannot be directly utilized by cells.

glucose: colorless crystalline or white crystalline or granular powder; It is odorless, sweet and hygroscopic. Soluble in water, easy to decompose when heated under alkaline conditions. It should be stored tightly closed.

After oral administration, it is quickly absorbed, and after entering the human body, it is used by tissues, and can also be converted into glycogen or fat storage. The normal body's ability to utilize glucose per minute is 6 milligrams per kilogram of body weight. It is a carbohydrate that can be directly absorbed and utilized, supplementing caloric energy, and is the main energy required by the human body, which is oxidized into carbon dioxide and water in the body, and at the same time provides heat, or is stored in the form of glycogen.

It can promote the detoxification function of the liver and have a protective effect on the liver. It is the most common energy material in living organisms.

Glucose is also known as dextrose. A monosaccharide, hexose containing aldehyde groups. The solid state is white crystal, slightly sweet when dissolved in water, and has optical rotation, and its aqueous solution is optical rotation to the right.

It is widely found in living organisms and is a component of certain disaccharides (such as sucrose, maltose, etc.) and polysaccharides (such as starch, cellulose, etc.). Free glucose is found in the fruits of certain plants (such as grapes) and in the blood of animals and is the main energy substance in living organisms. The glucose (blood sugar) content in the human blood is relatively constant, and the blood glucose concentration in a normal person is 80 120 mg when fasting and at rest.

It is an energy substance that is utilized by cellular respiration and does not provide nutrients. Don't forget the rewards uh.

As the embryo continues to develop, the required conditions are getting higher and higher, and the current level of human science and technology is still difficult to simulate the environment required for the fetus to develop in the mother's body.

Does not contain glucose, instead of lactic acid and/or pyruvate as energy**.

There are four types of animal cell culture media: balanced salt solution, natural medium, synthetic medium, and serum-free culture medium.

At present, there are two commonly used balanced salt solutions: sodium lactate and compound sodium chloride solution (the ratio of sodium lactate solution and compound sodium chloride solution is 1:2) and sodium bicarbonate and isotonic saline solution (the ratio of sodium bicarbonate solution and isotonic saline is 1:2).

Natural media, also known as compound media, are natural organic substances that contain unclear or unconstant chemical composition.

Natural medium is mainly extracted from automatic body fluids or isolated from animal tissues. Its advantage is that it is rich in nutrients and has a good cultivation effect, but the disadvantage is that the composition is complex and limited.

Synthetic medium, also known as combination medium, is prepared by sequentially adding accurately weighed high-purity chemical reagents with distilled water, and the components (including trace elements) and their amounts are known exactly. Synthetic media are generally used in laboratories for studies with high quantitative requirements, such as nutrition, metabolism, genetics, identification, and bioassays.

Serum-free culture media is a cell culture medium that does not contain serum but contains a variety of nutrients (such as growth factors, tissue extracts, etc.) that support cell proliferation and biological reactions.

Serum-free culture medium consists of basal culture medium and additional components that replace the serum supplement:

1. Hormones and growth factors;

2. Binding proteins;

3. Low molecular weight nutritional factors;

4. Adhesion and spreading factors, etc.

1.Amino acid.

The basic units that make up proteins. Different types of cells have different requirements for amino acids, but there are several amino acids that cannot be synthesized by cells themselves and must be provided by the culture medium, which are called essential amino acids. Among them, glutamine is an essential amino acid for cells to synthesize nucleic acids and proteins, and when glutamine is lacking, cells grow poorly and die.

2.Carbohydrates.

Carbohydrates are the main energy source for cell growth**, and some of them are components for the synthesis of proteins and nucleic acids. The main ones are glucose, ribose, deoxyribose, sodium pyruvate and acetic acid.

3.Vitamin.

Many vitamins are involved in the components that make up the active groups of various enzymes, without which the enzyme is inactive and metabolic activities cannot be carried out.

VA is the carrier of oligosaccharide groups when cells synthesize glycoproteins, and has an important maintenance role in epithelial cells. VD is involved in regulating calcium absorption. VE is an antioxidant that prevents the oxidation of unsaturated fatty acids in phospholipids, which make up biofilms.

VK deficiency causes hypothrombin and prolonged clotting time.

4.Inorganic salt ions.

Basic inorganic ions such as sodium, potassium, magnesium, calcium, phosphorus, etc., which are necessary for the composition of cells and participate in cell metabolism.

5.Animal serum.

Almost all animal cells grow and multiply in vitro in serum.

6.Nucleic acid. Nucleic acids are the basic building blocks of DNA and RNA, and basically animal cells can synthesize nucleic acids themselves, but most animal cells will be added to the in vitro culture.

7.Antibiotic.

Since it is inevitable that the cell culture in vitro will cause bacterial infection and cell death, an appropriate amount of antibiotics will be added to the culture medium.

MS medium.

Its composition is to add grams of ammonium nitrate, grams of potassium nitrate, grams of calcium chloride, grams of magnesium sulfate, grams of potassium dibasic phosphate, grams of potassium iodide, milligrams of boric acid, milligrams of magnesium sulfate, milligrams of zinc sulfate, milligrams of sodium molybdate, milligrams of copper sulfate, milligrams of cobalt chloride, milligrams of iron sulfate, 30 grams of sucrose and 7 grams of agar when preparing 1 liter (1000 ml) of medium. Other growth regulators should be determined according to the type of flower and the purpose of cultivation. The concentration of a large number of elements in MS medium is too high, so 1 2 and ?

A large number of element concentrations are used for cultivation, so that the growth effect is listed.

Before preparing the culture medium, prepare triangular bottles, test tubes, beakers, measuring cylinders, suction and other glassware, and weigh the medicine in advance. When preparing, the agar is dissolved first, and then various nutrients and sucrose are added to the deep hydrolysis in water, and then the pH of the medium is adjusted with sodium hydroxide or hydrochloric acid, which is generally mastered. Later, it can be dispensed into the bottle and capped.

The medium should be autoclaved. After cooling, put it in the culture room for 3 days of pre-culture, if there is no contamination with miscellaneous bacteria, the inoculation of flower materials can be carried out.

It depends on what you're used for. If the number of animal cells is simply cultured, then the composition of the medium of animal cells is: water, inorganic salts, animal serum, vitamins, sugars, nucleotides and various organic substances.

The medium generally contains carbohydrates, nitrogenous substances, and inorganic salts (including trace elements).

as well as vitamins and water, among others. Some media also contain antibiotics and pigments. According to the different raw materials used, it can be divided into two categories:

Those prepared with natural ingredients such as broth and potato juice are called natural mediums; The application of chemicals to prepare and label the composition is called synthetic medium or comprehensive medium.

Serum, glucose, amino acids, inorganic salts, vitamins.

Plant tissues.

The composition of the medium is mainly water, sucrose, mineral nutrients - inorganic salts, plant hormones and agar, etc. Edition. It is usually a solid-weight medium.

Animal cell culture medium is a liquid medium, the main components are water, carbohydrate-glucose, inorganic salts, growth factors and animal serum, etc. Animal serum can provide essential nutrients that are not yet known ... In addition, the composition and concentration of their inorganic salts are different...

Animals are generally added with serum, and plants are added with plant hormones.

Plant. Hormones in animals. Serum,