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Q：Should DMSO be removed immediately when thawing and culturing cells in cryovials?
Except for a few cells specifically marked as sensitive to DMSO, the vast majority of cell lines (including suspended cells) should be placed directly into a culture flask containing 10-15 ml of fresh culture medium after thawing. The fresh culture medium should be replaced the next day to remove DMSO, which can avoid the problem of most thawed cells being unable to grow or adhere.
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Q：How should the freezer be thawed?
After removing the freezing tube, it should be immediately placed in a 37 ℃ water tank for rapid thawing. Gently shake the freezing tube to melt it completely within 1 minute, and be careful not to let the water surface exceed the edge of the freezing tube cover, otherwise contamination may occur. When taking out the freezing tube from the liquid nitrogen tank for thawing, safety must be taken into account to prevent the freezing tube from bursting.
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Q：How should suspended cells be passaged?
Generally, it is only necessary to continuously add fresh culture based on the original culture flask and dilute the cell concentration. If there is too much culture medium, the mouth of the culture flask can be slightly raised until it cannot accommodate it. When dividing the bottles, take out a portion of the culture medium containing cells and transfer it to a new culture flask. Add fresh culture medium and dilute to the appropriate concentration. Repeat the above steps.
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Q：What is the seeding density of cells?
Inoculate according to the seeding density or dilution ratio on the basic data of the cell line. Too few cells or too many dilutions are also important reasons for the inability of cells to grow.
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Q：What do FBS, FCS, CS, HS abbreviations mean?
FBS (fetal bovine serum) and FCS (fetal calf serum) have the same meaning, both referring to fetal bovine serum. FCS is an incorrect term, please do not use it again. CS (calf serum) refers to calf serum. HS (horse serum) refers to horse serum.
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Q：How to choose a special cell line culture medium?
There are no fixed culture conditions for cultivating a certain type of cell. Cells cultured in MEM are likely to grow easily in DMEM or M199. In summary, choosing MEM for adherent cell culture and RPMI-1640 for suspension cell culture is a good start.
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Q：Where can I sell fetal bovine serum FBS for scientific research?
It is sold on the official website of Huayuan Liji Biotechnology. Each batch of raw blood materials for Liji fetal bovine serum has undergone strict screening and has been filtered through three consecutive layers of 0.1 μ m filter membranes. The filter material meets the requirements of the US FDA documents. The entire production process complies with international fetal bovine serum production standards and undergoes strict quality control and analysis and testing of physical, chemical, and microbiological indicators. Among them, serum contains proteins, amino acids, glucose, hormones, etc., among which proteins are mainly albumin and globulin. There are various types of amino acids, which are the basic components of protein synthesis in cells. Some of these amino acids cannot be synthesized by animal cells themselves (known as essential amino acids). The hormones contained in serum include insulin, growth hormone, and various growth factors (such as epidermal growth factor, fibroblast growth factor, insulin-like growth factor, etc.). Serum also contains various unknown factors that promote cell growth, adhesion, and other active substances, which can promote cell growth, proliferation, and adhesion.
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Q：What concentration of CO2 should be used when cultivating cells? 5% or 10%, or no impact at all?
Most culture media use HCO3-/CO32-/H+as a pH buffer system, and the content of NaHCO3 in the medium will determine the CO2 concentration that should be used during cell culture. When the NaHCO3 content in the culture medium is 3.7g per liter, 10% CO2 should be used for cell culture; When NaHCO3 is 1.5g per liter in the culture medium, cells should be cultured with 5% CO2.
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Q：Why does the purchased cell freezing tube have too few cells after thawing?
Researchers found that the number of cells after thawing was too low, mostly due to errors in the centrifugation process, resulting in physical damage to the cells and cell loss. It is recommended not to centrifuge the cells immediately after thawing. Wait for the cells to grow overnight before changing the culture medium.
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Q：How to avoid cell contamination?
The types of cell contamination can be divided into bacteria, yeast, mold, virus, and mycoplasma. The main reasons for cell contamination include improper aseptic operation techniques, poor operating room environment, contamination of experimental consumables, serum contamination, and contamination of cell sources. Strict aseptic operation techniques, clean environment, and high-quality cell sources are the best ways to avoid contamination.
"Heyuan Liji" is a joint venture between Heyuan Biotechnology (stock code: 688238) and Liji Biotechnology, specializing in the "Liji Biotechnology" and "Life-ilab" reagent brands
