What is cell culture?
What is cell culture?Cell culture is a modern biological technique that grows eukaryotic or prokaryotic cells in a controlled state. The development and methods of this technology are closely related to tissue culture or organ culture. The routine steps of cell culture include thawing cells, changing plates, dividing plates, changing cell culture medium, freezing cells, and the like.
Equipment and facilities for cell culture
Facilities: ultra-clean bench, constant temperature incubator, inverted microscope, liquid nitrogen storage tank, electric heating blast drying box, freezer, electronic balance, constant temperature water bath, centrifuge, pressure steam sterilizer.
equipment:
1. Glass equipment
Petri dishes, drip bottles, graduated pipettes, centrifuge tubes, culture flasks, beakers, measuring cylinders, conical flasks
2. Plastic equipment
Multiwell plates, dishes, flasks
3. Rubber equipment
Rubber products (preferably silicon products) are used as stoppers and lids for various bottles or test tubes.
4. Metal equipment
Scissors, forceps, scalpel, scalpel, vascular forceps, tissue forceps, ophthalmic forceps and various types of needles
5. Other items
Gauze, Syringes and Needles
The process of cell culture
1. Preparation
Preparatory work is extremely important to carry out cell culture, and the workload is also large, so it should be given enough attention. Negligence in a certain link in the preparatory work may lead to the failure or failure of the experiment. The preparatory work includes cleaning, drying and disinfection of utensils, preparation, sub-packaging and sterilization of culture medium and other reagents, cleaning and disinfection of sterile rooms or ultra-clean benches, inspection and debugging of incubators and other instruments. The content can be found in the relevant literature.
2. The material
A certain tissue cell is taken out from the body in a sterile environment (depending on the purpose of the experiment), and after a certain treatment (such as digestion and dispersion of cells, separation, etc.), it is inserted into a culture vessel. In the case of the expanded culture of cell lines, there is no process of taking materials. The first culture of tissue cells removed from the body is called primary culture. In theory, all tissues in various animals and humans can be used for culture. In fact, larval tissues (especially embryonic tissues) are easier to culture than adult tissues, and tissues with low degrees of differentiation are easier to culture than those with high degrees of differentiation. Tumor tissue Easier to culture than normal tissue. The tissue should be processed immediately after collection and cultured as soon as possible. If the tissue cannot be cultured immediately for some reason, the tissue block can be cut into small pieces as large as soybeans and stored in the culture medium at 4°C. Strictly maintain sterility when taking the tissue, and avoid contact with other harmful substances. Pathological tissue and skin and digestive tract epithelial cells are easy to carry bacteria, and antibiotics can be used to reduce pollution. Methods for dispersing cells from tissue and isolation can be found in the literature.
3. Cultivation
The process of transferring the obtained tissue cells into a culture flask or plate is called culturing. In the case of tissue block culture, directly connect the tissue block to the bottom of the culture vessel. After a few hours, the tissue block can be firmly attached to the bottom, and then add the medium. For cell culture, the cells should generally be counted before inserting into the culture vessel, and a certain amount (expressed as the number of cells per milliliter) should be inserted into the culture vessel as required, and the medium should be added directly. Immediately after the cells enter the culture vessel, they are placed in the incubator to allow the cells to enter the growth state as soon as possible. The cells in culture should be observed at regular intervals, including whether the cells are growing well, whether the morphology is normal, whether there is contamination, whether the pH of the medium is too acidic or too alkaline (indicated by phenol red indicator), and in addition The incubation temperature and CO2 concentration should also be checked regularly. Generally, there is a period of incubation period (ranging from several hours to tens of days) after the primary culture enters the culture. During the incubation period, the cells generally do not divide, but can adhere and migrate. After the incubation period, the cells enter the vigorous division and growth phase. After the cells have grown to the bottom of the flask, subculture should be carried out. The cells in one flask are digested and suspended, and then divided into two to three flasks for continued cultivation. Each passage is called a "generation". Diploid cells can generally only be passed down for dozens of generations, while transformed cell lines or cell lines can be passed down indefinitely. Transformed cells may be malignant in nature, or they may be immortality without malignancy. Cultivating growing cells is a good material for various biomedical experiments.
4. Cryopreservation and Recovery
To preserve cells, especially mutant cells or cell lines that are not readily available, cells are cryopreserved. The temperature of cryopreservation is generally the temperature of liquid nitrogen - 196 ° C. The cells are collected into a cryopreservation tube, and a medium containing a protective agent (usually dimethyl sulfoxide or glycerol) is added, and frozen at a certain cooling rate. Store in liquid nitrogen. At extremely low temperatures, cells can be kept for almost unlimited time. The recovery method generally adopts the quick-thaw method, that is, after taking out the cryovial from the liquid nitrogen, immediately put it into 37 ℃ water, so that it can be rapidly thawed within one minute. The cells are then transferred to culture vessels for culture. The selection of protective agent, cell density, cooling rate, temperature during recovery, and thawing rate during cryopreservation all have an impact on cell viability.
Laboratory consumables supplier:
Geneture medical specialized in the field of nucleic acid extraction system, PCR detection system and laboratory consumables, main products involved in：
1.Auto nucleic Acid Extractor （32T, 96T）
2.PCR fluorescence quantitative analyzer (16T, 96T)
3.Nucleic Acid Extraction Kit (Magnetic beads method)
4.Lab consumables: deep well plate, mag-rod sleeve, pipette tips, VTM, swabs, PCR tubes, PCR plate, centrifuge tubes, cryogenic Vials, saliva collection, cell culture dish/flask/plate/tube and so on.
Please don’t hesitate to contact with us to get a quotation.