LUOYANG FUDAU BIOTECH CO.LTD.--Cell Culture Consumables

The   cell factory   is a consumable that is often used for adherent cell culture. During the experiment, we need to grasp the growth state of the cells in real time and make relevant adjustments according to the specific situation. Cell viability can be detected by MTT method to determine cell growth. MTT method, also known as MTT colorimetric method, is a method for detecting cell survival and growth. The detection principle is that succinate dehydrogenase in the mitochondria of living cells can reduce exogenous MTT to water-insoluble blue-purple crystalline formazan and deposit in cells, while dead cells have no such function. Dimethyl sulfoxide (DMSO) can dissolve formazan in cells, and its light absorption value can be measured by enzyme-linked immunosorbent assay at 570nm wavelength, which can indirectly reflect the number of living cells. Within a certain cell number range, the amount of MTT crystal formation is proportional to the cell number. This method has been widely used i

Cell factories   occupy an important position in large-scale cell culture, mainly used in pharmaceuticals, vaccine production, cell therapy and other fields. Cells are more sensitive to the production environment, so the requirements for consumables are also relatively high. Biological detection is a very important basic detection, mainly including the following items: Cytotoxicity test: Pure cell-killing events caused by cells or chemicals that do not rely on apoptosis or necrosis as cell death mechanisms. Through this test, it can be judged whether the raw materials of the cell factory contain substances that are not conducive to cell growth. Sensitization test: chemical sensitizers act on the body through various ways, and can make immune cells in the body produce specific immune responses to them. damage, showing obvious symptoms and signs. Through this test, it can be judged whether the test substance is an allergen and the sensitization intensity. Hemolysis test: Hemolysis test i

  Nanobacteria is often encountered in cell culture. After the cells are contaminated, the adherent growth will not be affected in a short period of time, and the medium will not quickly turn yellow or turbid like bacteria and fungi. So, what to do if there is Nanobacteria  contamination in the  cell factory ? There is no clear conclusion about Nanobacteria, and it is generally believed that the black matter is the debris produced by the deterioration and death of cells due to various reasons. It can penetrate the filter membrane or spread through the air. It is black dots at low magnification, and black bugs can be seen swimming around at high magnification. The culture medium is also not muddy, generally it will not be too affected, and the cells can still be used. Usually, the cells grow well, and the observed movement does not increase significantly, and there is no obvious change in the color and transparency of the culture medium. Similar phenomena can be found in the same batch

Mycoplasma contamination is a very common problem when culturing cells in   cell factories . Unlike other contaminations, mycoplasma-contaminated cells generally do not become cloudy, so it is difficult to judge with the naked eye. There are four ways to determine whether cells are contaminated with mycoplasma: 1. DNA fluorescence staining The DNA fluorescence staining method is based on the principle that the fluorescent dye Hoechst33258 can bind to the AT base-rich region in the DNA of mycoplasma. Fluorescent dots are mycoplasma DNA rich in AT base regions. 2. PCR technology The specific primers are designed according to the conserved sequences in the Mycoplasma genome, the nucleic acid of the sample to be tested is amplified, and the diagnosis is made by analyzing the size of the amplified product. PCR detection technology is used for the detection of mycoplasma contamination, with short cycle, high sensitivity, good specificity, simple operation, and can detect a large number of sa

Mycoplasma contamination is a frequently encountered problem in basic research and industrial production. When using   cell factories   for cell culture or large-scale industrial production, mycoplasma contamination is also very troublesome. So, what to do in this situation? Mycoplasma is a microorganism whose size is between bacteria and virus (minimum diameter 0.2um) and lives independently. After mycoplasma contaminates cells, the culture medium may not become cloudy. It is relieved by passage and medium exchange, so it is easy to be ignored. But some serious cases can cause slow cell proliferation and even fall off from the cell factory. If mycoplasma contamination occurs, for non-important cells, such as cells in culture, cells of WCB, etc., the culture and the used medium can be inactivated and discarded. For more important cells, MRA treatment, drug-assisted warming treatment, use of mycoplasma-specific serum, inoculation and sterilization in animals, macrophage phagocytosis, an

  Bacterial contamination is a common source of contamination when using   cell factories   to culture cells, including Escherichia coli, Staphylococcus albus, Pseudomonas, etc. What should be done if the cells are contaminated by bacteria? Judgment of pollution: Bacterial pollution is different from other pollution sources. Once the cells in the cell factory are polluted, it is easy to detect. In most cases, the culture solution turns yellow in a short time, indicating that a large amount of acidic substances are produced, and obvious turbidity occurs; The culture medium was not mixed at first, but after a little shaking, many turbid substances floated. Observed under an inverted microscope, it can be seen that there are a large number of spherical particles floating in the culture medium, and sometimes a large number of bacteria exist on and around the cell surface, and the cells stop growing and show poisoning. When necessary, a small amount of culture medium smear can be taken for

Mold contamination is one of the things that researchers are very worried about in cell culture experiments, often leading to project delays and the loss of scarce cells. So how should mold contamination in   cell factories   be dealt with to reduce losses? Judgment of contamination: There are many types of fungi, most of which contaminate cells are Aspergillus, Candida albicans, yeast, black mold, spore fungi, etc. After mold contamination, light yellow or white floating objects are often formed in the culture medium, which are visible to the naked eye and can be easily detected. The culture medium generally remains cloudy in a short period of time. Under an inverted microscope, filamentous, dendritic or tubular hyphae that crisscross the cells can be seen, floating in the culture medium. Many hyphae can be seen in a chain-like arrangement of strains under high magnification. Candida and yeast strains are oval in shape and grow scattered on and around cells. Sometimes hyphae growing o

Cell factories   play an important role in large-scale cell culture such as vaccine preparation and lentiviral vectors. When culturing cells, various kinds of pollution are very troublesome. Among the pollution sources, microbial pollution is a relatively common type of pollution. Microbial contamination refers to various sources of contamination such as bacteria, molds, mycoplasmas, and black worms that grow when cells grow in cell factories. 1. Bacteria: Bacterial contamination commonly includes Escherichia coli, Staphylococcus, etc. This kind of contamination is easy to find. It is black and fine sand under an ordinary inverted microscope. The culture solution generally turns yellow in a short time. The liquid is not mixed, a little shaking will cause a lot of turbidity to float. 2. Mold: Most of the mold contamination is Candida albicans, Aspergillus, yeast and so on. After mold contamination, the culture medium remained clear and turbid in a short period of time. Under an inverted

Cell contamination refers to foreign substances that are mixed into the cell culture environment to produce harmful components and cause cell impurity. There are many common contaminations when culturing cells in   cell factories , among which chemical contamination is a very important and common type. Many chemicals in the culture environment can cause contamination of cells. Nor do these chemicals always inhibit cell growth. Some substances, such as hormones, can promote cell growth in cell factories, but unpurified substances, media, water, serum, growth cofactors, and containers for storing reagents can all become source of chemical pollution. 1. Essential nutrients for cell culture, such as amino acids, will also be toxic to cells if the concentration exceeds the appropriate range. 2. Different cell lines have different requirements for serum and buffer under the optimal culture conditions, which should be strictly controlled during culture. 3. The most common chemical contaminati

  Cell contamination is a very troublesome problem when culturing cells. Once the cells are contaminated, the cells will grow slowly, and in severe cases, they will lead to cell death and affect the experimental process. Physical contamination is a common type of contamination when culturing cells in   cell factories . Physical pollution mainly refers to the destruction of cells by physical factors such as temperature, vibration, radiation, and radiation. The optimum temperature for most cells to be cultured in vitro is 37-38°C. At low temperatures, the metabolic activity and mitotic capacity of the cells are reduced. If the temperature is not lower than 0 °C, although cell metabolism is affected, there is no damage; at 25 to 35 °C, cells grow at a slow rate; but if it is placed at 40 °C for several hours, it is not only unfavorable for cell survival and growth. , and can even lead to cell death within the cell factory. At the same time, the medium, buffer, fetal bovine serum, etc. use

When culturing cells, we are often troubled by various cell contaminations. Once the cells are contaminated, it will not only affect the cell growth, but in some cases, we have to discard the cells and re-cultivate them, which will affect the experimental process.   Cell factories   are widely used in large-scale cell culture, and their common contaminations mainly fall into the following three categories: 1. Physical pollution: mainly refers to the damage to cells by physical factors such as temperature, vibration, radiation, and radiation. Such as exposure of cell culture medium to radiation or ultraviolet light, it will cause changes in cell metabolism. There are devices around the incubator that can generate mechanical vibrations, which may also have a certain impact on cell growth. 2. Chemical pollution The medium and water used for cell culture should be sterilized in an autoclave. Among them, serum is a commonly used medium in cell culture, and serum has potential chemical conta

When culturing various cells,   cell culture flasks   are a kind of cell culture consumables that are used more frequently. They are mostly used for the culture of adherent cells, and the specifications range from 25ml to 225ml. The hamster kidney cell culture medium will use this consumable, and the specific steps of its culture are as follows: 1. Preparatory work for cell culture: 1) Prepare MEM medium; high-quality fetal bovine serum, 10%; double antibody 1%. 2) Culture conditions: Gas phase: air, 95%; carbon dioxide, 5%. Temperature: 37 degrees Celsius, incubator humidity of 70%-80%. 3) Freezing solution: 90% serum, 10% DMSO, ready-to-use. 2. Cell processing: 1) Recovery of cryopreserved cells: Quickly shake and thaw the cryopreserved tube containing 1 mL of cell suspension in a 37°C water bath, add it to a centrifuge tube containing 4-6 mL of complete medium and mix well. Centrifuge at 1000RPM for 3-5min, discard the supernatant, and resuspend the cells in complete medium. The cel

The   cell factory   is a multi-layer structure of cell culture consumables. Common specifications include 1 layer, 2 layers, 5 layers, 10 layers, 40 layers, etc. The higher the number of layers, the larger the culture area. This consumable is mainly used for the culture of adherent cells, where the following conditions are necessary for cell growth: 1.  S terile  e nvironment Nontoxicity and sterility are important conditions for culturing cells in cell factories. In vivo, the detoxification system and immune system can resist the invasion of microorganisms or other harmful substances, but in the process of in vitro culture, cells lack the protection of the body's immune system and lose the ability to defend against microorganisms and detoxify harmful substances. To ensure that cells can grow and reproduce in an in vitro environment, it is necessary to ensure a sterile work area, good personal hygiene, sterile reagents and media, and aseptic handling. 2. The right temperature Gene

  Cell factory   culture technology plays an important role in large-scale cell culture such as vaccine development and biopharmaceuticals. Various solutions are needed for cell culture, and PBS buffer is one of them. PBS is the English abbreviation of Phosphate Buffered Saline Solution. Generally, active biological preparations are diluted with it, and pseudorabies virus can also be diluted with it. This is because the PBS buffer has an adjustable pH buffering effect and salt balancing effect. The reason for not using distilled water is that water destroys the structure and biological properties of biological proteins; the reason for not using normal saline is that it cannot adjust the pH. Therefore, the first choice for washing biological cells is PBS buffer. PBS is not a panacea. Maintaining optimal conditions ensures that biologically active substances maintain their most complete characteristics. Some biologically active substances require higher conditions than PBS. In this case,

As a common consumable for large-scale cell culture,   cell factories   play an important role in vaccine development, biopharmaceuticals and other fields. Cell culture is a special task. After all preparations are done, it is necessary to grasp the growth state of cells in time to make adjustments to each work. Depending on the culture area, the cell factory consists of one or more layers such as 1 layer, 2 layers, 5 layers, 10 layers, and 40 layers. Observing the growth state of cells generally requires an inverted microscope, but this instrument usually only It is suitable for observing 1-10 layers of cell factories, and can observe the growth of 1 and 2 layers. The bottom second layer can only observe the uniformity of its cells, and it is not suitable for forty layers. If you want to observe the growth state of cells in more layers of cell factories, you need to use microscopes specially designed to observe cell factories. Such instruments generally have lenses in two directions:

Cell Factory   is a multi-layered cell culture consumable suitable for large-scale cell culture. In addition to a specific environment for cell culture, digestive juice is also an essential solution. In the in vitro culture of tissue cells and the dispersion of tissue cells in primary cell culture (the tissue block is prepared into a single cell suspension) as well as in subculture, the digestion and dispersion of adherent growing cells should use tissue cell digestion solution. The digestion solution used in cell factory culture technology is mainly trypsin, EDTA, etc. The main components of the cell membrane and the adhesion between cells are lipoproteins and polysaccharides. Trypsin in trypsin can convert proteins into peptones, and pancreatic lipase can decompose fat into glycerol and fatty acids, thereby separating cells. Cells are lysed. The function of the digestion solution is mainly to hydrolyze the intercellular proteins (such as extracellular matrix), to disperse the tissue

As a cell culture consumable,   cell factory   has become the main consumable for large-scale cell culture such as pharmaceutical companies and vaccine production. Cell culture requires specific conditions, of which a sterile environment is the primary condition for in vitro cell culture. In vivo, the detoxification system and immune system can resist the invasion of microorganisms or other harmful substances, but in the process of in vitro culture, cells lack the protection of the body's immune system and lose the ability to defend against microorganisms and detoxify harmful substances. To ensure that cells can grow and reproduce in an in vitro environment, it is necessary to ensure a sterile work area, good personal hygiene, sterile reagents and media, and aseptic handling. Including the cell factories used are all produced in C-level clean workshops, and they must be sterilized after production to ensure that the consumables themselves have no factors that hinder the growth of c

The development of cell culture technology has been very mature so far, and it has been widely used in the preparation of monoclonal antibodies, vaccine production, scientific research and other fields. Cell culture requires the help of cell consumables. There are various types of consumables on the market. Among them, the   cell factory   is a kind of consumable with special structure. Different from other bottle-type consumables, Cell Factory is a special consumable with a multi-layer structure. Common specifications include 1 layer, 2 layers, 5 layers, 10 layers, 40 layers, etc. When culturing cells, the medium is poured into the inner layers, and the cells are attached to the bottom to grow and multiply. Due to the high number of 10-layer and 40-layer layers, it is difficult to operate after adding the medium. Generally, it is used together with an automated shaker. It can realize the programming, automation and efficiency of large-scale cell culture, thereby Greatly reduce labor i

Measles vaccine is a combined live attenuated vaccine that stimulates immunity against measles virus, mumps virus and rubella virus after vaccination. It is used to prevent measles, mumps and rubella. In the large-scale production of this vaccine, the   cell factory   is an indispensable cell culture consumable. The measles vaccine is to inoculate chicken embryo cells with attenuated strains of measles virus and attenuated mumps virus respectively, and inoculate human diploid cells with attenuated strains of rubella virus. Add suitable stabilizer and freeze-dried. It is a cheese-colored loose body, which is orange-red or light pink clear liquid after reconstitution. The main components of lyoprotectant are human albumin, gelatin and sucrose. In the past two years, the incidence of measles in my country has shown a downward trend. In 2019, the number of measles cases in my country was 2,974, the number of deaths was 0, and the incidence rate dropped to 0.21/100,000. In 2020, the cumulat