Cell culture is both complex and complex, simple and simple, the maintenance of the cell culture environment, the selection of culture media and the recovery of cells are all crucial, today to share what basic conditions are needed for cell culture.
Suitable cell culture media
Culture medium is a solution that maintains cell survival and growth in vitro and is divided into natural medium and synthetic medium. Now the laboratory generally uses synthetic medium, synthetic medium now has DMEM, 1640, IMDM, L15, etc., different cells to use different kinds of medium, such as adherent cells can be mostly cultured with DMEM, suspension cells are mostly cultured with 1640.
The quality of the serum
Synthetic media can only maintain cell survival, and in order for cells to grow and reproduce, high-quality serum needs to be supplemented to increase nutrients. Serum is one of the most important components of culture media, containing a variety of growth factors and nutrients required for cell growth.
Sterile, non-toxic cell culture environment
The sterile and non-toxic culture environment is the primary condition to ensure the survival of cultured cells. The human body has a strong immune system and detoxification organs, so when toxic substances invade the body, organs such as the immune system and liver can resist and remove them. However, cells lose their resistance when cultured in vitro, and once contaminated cells will die, so maintaining a sterile and non-toxic environment is the most basic condition for in vitro cultured cells.
Constant cell growth temperature
To maintain cell proliferation and growth, it is necessary to have the right temperature. At present, most of the cells cultured in the laboratory are mainly mammalian cells such as human, rat, mouse, pig, etc., and the general cell culture temperature is about 37, at which the cells can grow and metabolize normally. When the temperature is high, higher than 39 or higher can cause damage or even death to the cellsWhen the temperature is low, it is not lower than 0, although it will affect the metabolism and growth state of the cell, it will not cause the cell to die, and most of the cells can recover their growth characteristics after the temperature is restored. Very few laboratories have insect cells such as SF9, Hi-Five, etc., which need to be cultured in an environment of 28, and the specific culture temperature must pay attention to the cell bank and the corresponding manual information.
Suitable gas atmosphere
The appropriate gas environment is also one of the conditions for cell survival, the required gases are O and Co, O to produce energy to make cells grow and proliferate and synthesize various required components, Co is a cell metabolite and a component required by the cell, and its main role is to maintain the pH of the medium. Most media are filled with 95% air and 5% CO, but L15 medium, as a separately designed culture system, needs to be cultured directly under 100% air conditions, and the introduction of 5% carbon dioxide will cause the medium to be pH acidic and cause the cells to deteriorate or die.
Cell chamber disinfection is a very important task as it prevents contamination and cross-contamination in cell culture. Among the traditional cell room disinfection methods, the most commonly used are physical and chemical disinfection methods, and below we will compare the traditional cell room disinfection methods with the new environmentally friendly and safe disinfection methods of Inlink Life Sciences.
Physical disinfection methods
Physical disinfection is the method of killing or removing microorganisms by physical means. The most common physical disinfection methods are ultraviolet disinfection and high-temperature disinfection.
1. Ultraviolet disinfection.
UV disinfection is the use of ultraviolet light to kill or remove microorganisms. UV disinfection has many advantages, such as being fast, easy to operate, and does not produce chemical residues. However, UV disinfection also has some drawbacks.
First of all, it can only kill the surface of the microorganism, not the inside of the microorganism.
Second, UV light only kills microorganisms that are directly exposed to UV light, so every corner of the cell chamber needs to be exposed to UV light.
Finally, UV disinfection requires regular replacement of UV lamps, which can increase costs.
2. High temperature disinfection.
Hyperthermic disinfection is the killing or removal of microorganisms by high temperatures. High-temperature disinfection has many advantages, such as the ability to kill the interior of microorganisms and the absence of chemical residues. High-temperature disinfection has the following disadvantages:
First of all, high-temperature disinfection requires the use of high-temperature equipment, which increases costs.
Secondly, high-temperature disinfection takes a certain amount of time, so it needs to be planned and arranged.
Finally, autoclaving may cause damage to some cell cultures.
Chemical disinfection methods
Chemical disinfection is the method of killing or removing microorganisms through chemicals. The most common chemical disinfection methods are alcohol disinfection, hydrogen peroxide disinfection, and chlorine disinfection.
1. Alcohol disinfection.
Alcohol disinfection is the use of alcohol to kill or remove microorganisms. Alcohol disinfection has many advantages, such as being fast, easy to operate, etc. However, the biggest disadvantage of alcohol disinfection is that it cannot be used in a large area, and alcohol can only kill those microorganisms that come into direct contact with alcohol, so it is necessary to ensure that every corner of the cell chamber can be exposed to alcohol. In addition, alcohol disinfection may cause damage to some cell cultures.
2. Hydrogen peroxide disinfection.
Hydrogen peroxide disinfection is the killing or removal of microorganisms by hydrogen peroxide. Hydrogen peroxide disinfection is fast and does not produce chemical residues. But,Hydrogen peroxide disinfection has a corrosive risk, which is the biggest drawback, and all the choice of hydrogen peroxide disinfection method must consider whether the company has the ability to reduce the corrosive risk, such as equipment unit usage, corrosivity report, atomized particles, etc.
3. Chlorine disinfection.
Chlorine disinfection is the use of chlorine to kill or remove microorganisms. However, chlorine disinfection may produce harmful gases that need to be discharged.
In summary, traditional methods of cell compartment disinfection have their own advantages and disadvantages. When choosing a disinfection method, you need to choose according to the actual situation. For example, if rapid disinfection is required, UV disinfection or alcohol disinfection can be chosenIf thorough disinfection is required, high-temperature disinfection or chlorine disinfection can be chosen. When using chemical disinfection methods, it is necessary to pay attention to the concentration of chemicals and the contact time to avoid damage to cell cultures. At the same time, the performance of disinfection equipment needs to be checked regularly, and maintenance and replacement need to be carried out.
For cell culture laboratories, the most important thing is to eliminate all possible sources of contamination.
To solve and prevent environmental pollution, the cell room can use the space environment disinfection technology of Runlian Life Sciences: Norford Sporocide + Billingcohan hydrogen peroxide sterilization equipment.
Billingehan dry mist hydrogen peroxide sterilizer uses the world's most advanced atomization technologyThe atomized particle size reaches an average of 3-5 microns, with the use of Norford fungicide,The sterilization effect can reduce the spores of steatobacterium thermophilus by 6 logs。The atomization particles are small, which increases the uniformity of spatial diffusion, and quickly fills the entire confined space without letting go of every dead end, and its uniformity, no residue and no harm to the machine are very consistent with the disinfection of the cell room environment and the disinfection inside the incubator.
Norford sporicide is a product specially used to remove bacteria, orthobacteria, mycoplasma and other microorganisms in the cell room, and has been verified by a number of independent tests to show that Norford sporicide has a removal effect on all kinds of bacteria, orthobacteria, mycoplasma and other microorganisms in cell culture, especially for bacteria, fungi and their spores and spores exposed to the air can be inhibited for up to 7-10 days, which can be quickly removed. The main ingredient is high-purity hydrogen peroxide reactive adhesive paper silver ions, and its characteristic advantages include:
Broad-spectrum: it has a scavenging effect on viruses, bacteria, mycoplasma, fungi and molds
Safety: It has a third-party test report that is non-teratogenic, non-irritating and non-toxic, and it is safe to use
Targeted: Specially designed for use in cell rooms and laboratories, rapid removal of microbial contamination within 3-5 minutes:
Physical and chemical properties: non-irritating, non-corrosive, good stability, completely degradable;
Easy to use: It can be sprayed, atomized, fumigated, easy to operate, and there will be no problems such as bacterial resistance.
The intelligent hydrogen peroxide disinfection machine of Runlian Life Science is simple to operate, friendly to personnel, and can be used immediately after the equipment is powered on, starting and stopping at any time, and the sterilizer can complete the entire disinfection process outside the disinfection space, avoiding the normal time and emergency situation, the operator is exposed to the aerosol of the disinfectant solution in order to turn off the equipment, which is the preferred choice for the disinfection of laboratory and cell room space.
Specializing in solving the problems of intercellular mycoplasma pollution, fungal pollution, phage contamination and black gum insect pollution, we have rich experience in treatment, and can provide cell room disinfection and sterilization equipment, liquid, and door-to-door disinfection services and guidance, including Shanghai cell room door-to-door disinfection service, Suzhou laboratory door-to-door disinfection service, Nanjing laboratory door-to-door disinfection service, Beijing cell room door-to-door disinfection service, Guangzhou cell room pollution door-to-door disinfection, Shenzhen cell room door-to-door disinfection.