Balanced salt solutions. Hanks Saline Solution Hanks medium composition

Thalasso salt peeling Scrub based on natural marine ingredients. Promotes the removal of waste and toxins, stimulates blood circulation, cleanses and nourishes the skin. Thalasso peeling is carried out using marine products: salt, crushed seaweed,

• Immunization (lat. immunis free, free from something; synonym: immunoprophylaxis, protective vaccinations, preventive vaccinations) is the specific prevention of infectious diseases of people and animals. Immunoprophylaxis of a number of infectious diseases with airborne droplets...

News about Hanks saline solution

• G.V. Pavlov, N.V. Nikitina Scientific publication, Yekaterinburg, 2003 Introduction Many branches of theoretical and practical medicine, to one degree or another related to the functioning of the exocrine organs and tissues system in normal and pathological conditions, have reached high levels in recent decades.
• Corresponding member RAMS, Professor I.I. Balabolkin, Doctor of Medical Sciences L.S. Namazova, Ph.D. I.V. Sidorenko, professor I.S. Elkis, Ph.D. A.V. Topolyansky, professor A.L. Vertkin Scientific Center for Children's Health of the Russian Academy of Medical Sciences, Moscow MMA named after I.M. Sechenov MGMSU named after. ON THE. Semashko Emergency and Emergency Medicine Station

Hanks's discussion of saline solution

• I don't understand anything! I’ve been eating “isotonic” for a month now - minimum fat, sweets, balance of proteins and carbohydrates, and for the third week I’ve been doing shaping every other day. On the scales - as it was (from 58 to 60 with a height of 1.58 normosthenic build), on a centimeter - the same volumes, well, maybe a couple of centimeters have gone
• sodium hydrochloride Aqueous (0.9%) solution of sodium chloride. Plasma replacement agent. Its other name is isotonic sodium chloride solution for injection Pharmacology. Isotonic sodium chloride solution replenishes sodium deficiency

Cultivation of viruses in cell culture.

Cells obtained from various organs and tissues of humans, animals, birds or others biological objects, are capable of multiplying outside the body on artificial nutrient media in special laboratory containers (mattresses, bottles, test tubes, etc.). Cell cultures from embryonic and malignantly degenerated tissues, which have a more active ability compared to normal cells of an adult body, have become widespread. growth and reproduction.Depending on the preparation technique, three types of cell cultures are distinguished:

· single-layered - cells capable of attaching and multiplying on the surface of chemically neutral glass laboratory glassware in the form of a monolayer;

Suspension - cells that multiply throughout the entire volume nutrient medium with constant stirring;

Based on the number of viable generations, cell cultures are divided into:

· primary, capable of reproducing only in the first generations, i.e. in several passages after isolation from tissues;

· graftable, or stable, capable of reproducing in laboratory conditions indefinitely through constant passaging;

· semi-transplantable, having a limited lifespan (40 - 50 passages).

Animal and human cell cultures have certain requirements for the liquid (nutrient medium), gaseous (gas concentration) and solid (substrate surface) phase. When preparing nutrient media for cellular structures, one has to solve two difficultly compatible problems. On the one hand, it is necessary to reproduce an environment as similar as possible to the environment in which cells exist in vivo; on the other hand, in order to create controlled standard conditions, it is necessary to know the exact composition of the environment.

Natural or natural environments: Cavity fluids (amniotic or allantoic fluid, embryonic extract, tissue extracts, plasma or serum, GLA - lactalbumin hydrolysate - a product of enzymatic hydrolysis of milk) allow us to solve only the first problem. In media prepared from purified, strictly defined substances, natural conditions are only partially reproduced. These problems are easily solved during short-term cultivation, when the main role is played by factors such as osmotic pressure, pH, concentration of other inorganic ions, buffer capacity, and atmospheric composition. Osmotic pressure in a cell at 37°≈7.6 atm.

Changes in osmotic pressure within +-10% have almost no effect on the condition of the cells, and the pressure remains at a normal level, which is achieved by adding NaCl, taking into account other inorganic ions and glucose. It turns out that if we proceed from the need for stable osmotic pressure, it is easier to change the concentration of protein than of low molecular weight, especially inorganic compounds.

The optimum pH – 7.2 – 7.4 is controlled by a buffer system simulating the blood buffer system (NaHCO3 on a weak phosphate buffer).

Required for cell survival in vitro inorganic ions Na, Ca, Mn, Fe, CO2, PO4, SO4. Their role is not fully understood, but they are necessary to maintain osmotic pressure, cell attachment to glass, and enzyme activity. Therefore, the basis of any nutrient medium is balanced in osmotic pressure, pH, etc. saline solution

Earle's and Hanks' saline solutions are commonly used to prepare culture media. These solutions, like Dulbecco's and Vogt's phosphate buffered saline, are also used for irrigating and washing cells during passage of cultures, isolation of cell lines and other manipulations with cell cultures.

Earle and Hanks salt solutions with a high buffer capacity of at least 3 ml (Hanks salt solution Ca Mg K Na - phosphorus, sulfur salts hydrochloric acids, D-glucose, phenol red.; Erla - poorer) + serum, amniotic fluid, embryonic extracts, yeast extract. The pH and osmolality ranges at which cell proliferation occurs are narrow and vary depending on the cell type. To maintain pH, most media use a bicarbonate buffer: HCO3 = CO2 + OH, if carbon dioxide is released, the OH concentration increases. Solutions may contain small amounts of bicarbonate buffer (Hank's solution) and are intended to maintain pH in tightly sealed containers. Others (Earl's solution) contain more bicarbonate and are used in systems with increased partial pressure of CO2. If cultures are maintained outside a CO2 incubator, where pH is more difficult to maintain, alternative buffer systems are needed. A good buffer is HEPES 4-(2-hydroxyethyl)1-piperazineethanesulfonic acid. Indicators are added to nutrient media in order to control the acidity of the medium at any given time. During growth, cells acidify the medium, and the indicator becomes yellow-orange in color.

Synthetic, standard media: medium 199, Eagle medium and its modifications, RPMI1-1640 medium, they contain many components necessary for cell development: amino acids (10 essential ones as well as cystine and tyrosine, many cells require glutamine), water-soluble vitamins, especially group B, synthesis coenzymes ATP, choline and inositol, acting as a hydrocarbon substrate. The media are based on Earle or Hanks solutions. They are used as supporting media for primary cells and for the cultivation of lymphocytes and hematopoietic cells.

Normal, maintaining specific functions cells on standard media do not reproduce (unless transformed). For optimal cell growth, 5-20% fetal serum is usually added.

Serum is an extremely complex mixture of small and large molecules that can both promote and inhibit cell growth. The main functions of serum include: providing hormonal factors that stimulate cell growth and their functions; providing factors for cell attachment and spreading; providing transport proteins that carry hormones, minerals, lipids, etc. Serum proteins directly and specifically involved in stimulating cell division are called growth factors.

Most growth factors are present in serum at concentrations of a few nanograms per milliliter or less. Some of these factors are specific to cells at a certain stage of differentiation; the action of others is not limited to any one type of cell. The same cell type can be stimulated by different growth factors. For example, fibroblasts proliferate in response to fibroblast growth factor, epidermal growth factor, platelet-derived growth factor, and somatomedins. All these substances are mitogens (stimulate mitosis). Another important growth factor for almost all cell types is the hormone insulin. Of the other hormones, the most commonly used are glucocorticoids (hydrocortisone, dexamethasone), steroids (estradiol, testosterone, progesterone) and thyroid hormones (triiodothyronine). Hormones stimulate or inhibit growth depending on the type of cells and their density. Glucocorticoids, for example, affect cell proliferation by changing their sensitivity to growth factors.

To transport low-molecular factors (vitamins, amino acids, lipids and others), transport proteins are required. Albumin plays this role. Iron transport is provided by transferrin, and the surface of most cultured cells contains receptors for this protein. The factors of cell attachment and spreading include collagen and fibronectin; chondronectin (chondrocyte adhesion) and laminin (epithelial cell adhesion) are more specialized.

Balanced salt solution (BSS) contains inorganic salts. In addition, it may contain sodium bicarbonate and, in some cases, glucose. The composition of some of the most used balanced salt solutions is presented in table. 9.2. Also, if necessary, HEPES buffer (5-20 mM) can be added to these solutions, while an equivalent amount of NaCl is removed to maintain osmotic pressure. Many complete environments and environment providers are based on BSS, which
Some people sell the medium MEM Needle with Hanks' salts or Eagle MEM with Earl's salts indicate which BSS composition was used in the preparation of the medium. Hanks' salts are used when culturing cells in closed flasks in an air atmosphere, while Earle's salts are used at a high concentration of bicarbonate in combination with 5% CO2 in the gas phase.

BSS is also used for diluting amino acid and vitamin concentrates in the preparation of culture media, for isotonic washes or preparations, and for short-term cell incubation (up to 4 hours, usually with the addition of glucose). The composition of BSS is often modified—for example, by eliminating glucose or phenol red from Hanks' BSS or eliminating Ca2+ and Mg2+ ions from Dulbecco's PBS. PBS without Ca2+ and Mg2+ is known as PBS A solution, and the designation D-PBSA will be used throughout this book to indicate the absence of these divalent cations. The modification must always be specified when purchasing BSS and in publications and reports.

The choice of BSS depends both on the partial pressure of CO2 (see Section 9.2.2 and Tables 9.1 and 9.2) and on the objectives of the researcher. If BSS is used as a tissue or cell monolayer disaggregation solution, Ca2+ and Mg2+ are usually excluded, such as in Moscona's calcium-magnesium-free saline (CMF) or D-PBSA (see Table 9.2). The choice of BSS also depends on whether the solution will be used for suspension culture or monolayer (attached) cell culture. Eagle's saline-based S-MEM, as modified by Spinner, is a variant of Eagle's minimal nutrient medium with a reduced Ca2+ content to reduce cell aggregation and cell attachment (see Table 9.2).

HBSS, EBSS and PBS as phosphate-based buffers have low buffering capacity at physiological pH. Paul proposed Tris-buffered BSS, which is more effective as a buffer, but requires adaptation for certain cell types. The most effective buffers are currently considered to be HEPES (10-20 mM) in the pH range 7.2-7.8 and Tricine in the pH range 7.4-8.0, but the use of these buffers on a large scale increases the cost of research significantly.