How To Store And Maintain Tissue Cultures

Certainly, you have heard about the ability of technology to grow cells and tissues in media then preserve them later on for future study or usage. It is a very important innovation as tissue cultures have been proven to be beneficial to mankind, particularly in diagnosing diseases and discovering cures. Concrete examples would be the development of vaccines and cures for certain diseases. However, it would be good to note that for tissue cultures to remain viable and usable, they should be properly stored and maintained.

Tissue cultures are appropriately maintained and grown at a temperature equivalent to the body core temperature which is 37 degrees centigrade. The gas mixture in the culture and around it should also be accurately measured like 5% of carbon dioxide. The preservation of the tissue culture is likewise critical, usually making use of liquid nitrogen in liquid or vapor phase. Since freezing can actually kill cells because of the extremely low temperature, chemical agents are added to lower the freezing point so that it would only freeze the liquid part of the culture but maintain the viability of the cells. Glycerol or DMSO are the usual agents used.

A simple device called the Mr. Frosty is used for the storage of tissue cultures. The Mr. Frosty is filled with isopropanol at 200 ml where the freezer vials are placed then it is placed in liquid nitrogen.

To be able to be sure that the cells and tissues remain viable, the cultures should be checked daily. Everything should be noted like the morphology, the density of the cells and the color of the medium. All notes should be put in a daily logbook that states the cell line, the components of the media and any changes made to the standard medium, dates of feeding, the computations for the culture doubling time and other changes noted.

To store the tissue culture, the use of medium that is expired can make the culture null and void. Media storage instructions should always be followed. If you see that the medium appear cloudy and with precipitates, dispose it immediately. The culture should be kept in a dark place and put in a refrigerator with a temperature of 2-8 degrees.

The reason why darkness is need is because exposure to light rays and temperature extremes can break down the components of the medium used. Also, high temperatures would allow the growth of by-products rendering the culture toxic. On the other hand, extremely low temperatures could cause precipitation in the medium that is irreversible.

In storage, it is also important to prevent any microbial contamination or even cross-contamination with other cultures. This could lead to cell and tissue death in the culture. Using aseptic technique in culturing and preservation with the use of Bio-safety cabinets can help in preventing the growth of microbes.

In maintaining the culture, here are the problems you should note to see if problems are already being encountered. To see if there is spotting, check for bubbles for these may have displaced some cells. If you see any streaking, there may be condensation buildup or an even an incubator problem. Uneven growth surfaces may mean clumping of cells where they were not totally suspended before seeding.

Other mistakes may be seen in stringy cells where there must have been overexposure to trypsin or pipettes have poor vacuums. A “halo” effect would mean poor distribution of the medium used over the cells, probably because of the flask design or unleveled incubator. If there is peeling, incubation may have been prolonged or there must have been overgrowth of the tissues cultured. Banding in the roller bottles may mean the rollers may have been turning too quickly or too slowly.

Added information on maintenance can be seen in the following. You should always use growing cells at the log phase of the growing stage. Make sure that they are already 80-90% viable. Use trypsin minimally. Cells should be handled also with much care, making sure that they are centrifuged at moderate speeds only. Medium to be used should also be serum-free.

Storing and the maintenance of tissue cultures is no joke. So many factors should be considered. Careful daily logging should be maintained to see if there are untoward incidences in the cultures. Do not forget that these cultures are also very sensitive but useful so it is important to know the proper way of storing and maintaining these cultures. Your cultures that are properly maintained may mean the life of a person, so make you sure you take extra care in handling those tissue cultures.