Consult on the culture of MDCK cells

Cell culture contamination is often the most common problem encountered in cell culture labs, and can sometimes have serious consequences. There are two main categories of cell culture contaminants, one is chemical contaminants such as: impurities in culture media, serum and water, endotoxins, plasticizers and detergents, and the other is biological contaminants such as:

Cross-contamination of bacteria, molds, yeast, viruses, mycoplasma, and other cell lines. Although contamination cannot be completely eliminated, the frequency and severity of contamination can be reduced by having a good understanding of the source of contamination and good aseptic technique. Bacteria are a large group of single-celled microorganisms that are widely present.

Bacteria are generally a few microns in diameter and come in a variety of shapes, such as spherical, rod-shaped, and spiral-shaped. Due to their wide distribution, rapid growth, and size, bacteria, along with yeast and mold, constitute the most commonly encountered contaminants in cell culture.

Once the culture is contaminated with bacteria, it can be detected by simple visual observation within a few days; Infected cultures are usually cloudy (i.e., cloudy) and sometimes covered with a thin film. In addition, it is not uncommon to see a sudden decrease in the pH of the medium.

Bacteria are tiny particles that move between cells under a low-power microscope, and the shape of each bacterium can be discerned when viewed under a high-power microscope. Yeast is a single-celled eukaryotic microorganism in the fungal kingdom that varies in size from a few microns (common) to 40 microns (rare). Similar to bacterial contamination, cultures can become cloudy after yeast contamination, especially in the later stages of contamination.

The pH of the culture changed very little when it was contaminated with yeast, and it only increased when the contamination was severe. Under the microscope, yeast appears as a single oval or spherical particle, with some budding to produce smaller particles. Mold is a eukaryotic microorganism in the fungal kingdom that grows in the form of multicellular filaments known as hyphae.

These multicellular filaments form a cross-linked network of genetically identical nuclei, known as colonies or mycelium. Similar to yeast contamination, the pH of the culture remains stable at the beginning of mold contamination and rises rapidly as the contamination increases, resulting in turbidity of the culture. Microscopically, mycelium usually appears as fine bundles of fibers, sometimes as dense clumps of spores.

The spores of many species of molds are resistant to extremely harsh, hostile environments during the dormant period and are activated only when they are exposed to the right growing conditions.

First, let's talk about MDCK cells.

MDCK was built in September 1958 and was developed in Cox's Banny kidney, which resembles fibroblasts in morphology during primary culture, and is purified into epithelioid cells by 6 consecutive digestions of trypsin and EDTA mixed digests, with an interval of 7 days between each purification. MDCK cells in culture have the differentiation characteristics of distal renal tubules and collecting ducts, ** cells. There are at least two cell types in the MDCK cell line, one named C5A, which secretes fluid, which leads to cyst formation, when grown in a culture containing type I collagen, but when cultured on a plastic-sided low material, it secretes fluid, but does not form a cyst, and the other cell is the opposite.

Uses of Cells:

MDCK cells are widely used as models for distal convoluted tubules or collecting ducts, and can also be used for metabolic studies and PG-level drug-drug interaction studies, as well as to observe the effects of influenza virus strains on cell function.

For cell culture, some do use DMEM medium, some use MEM medium, and there is no fixed culture condition for cultivating a certain type of cell. Cells cultured in MEM are likely to grow equally easily in media such as DMEM or M199. So I think you can choose as much as you want.

I used to prefer DMEM media.

4.The difference between high-sugar DMEM and low-sugar DMEM is:

The glucose content of high-glucose DMEM is, and the sugar content of low-sugar DMEM is, in which the sugar content of low-sugar DMEM is the same as that of general media, so it can be used with other general culture media, while high-sugar DMEM is mainly used in the culture of special cells such as tumors, and how to choose needs to be determined according to the requirements. I recommend that you use low glucose DMEM medium.

5.Fetal bovine serum is best used for serum, and calf serum can also be used if not. Fetal bovine serum** is a bit more expensive.