Pdf cell line crosscontamination and accidental coculture. An overview find, read and cite all the research you need. Tissue culture waste disposal guide updated may19 t. Customers in europe, africa and the middle east should contact our office in leiden, the netherlands. Cell culture is a fundamental technique for a variety of applications. Cross contamination of a cell culture with other cell types is a serious problem that has only recently been considered alarming. Today cell culture methods are vital to broad areas of life science research. The most frequent causes of cell culture contamination are. Elimination of mycoplasma contamination in cell cultures.
This is the first in a threepart series on co 2 incubation. How to prevent contamination with candida albicans during. Culture contaminants may be biological or chemical. If it is intended to genetically modify the cell culture in including the immortalisation of the anyway. However, 3070 days following cell culture contamination the increase in chromosomal aberrations was observed in the contaminated cell line primarily at the expense of chromosomal breaks. The method employs the following sequence of operations. The picture shows a petri dish with bacterial contamination. For animal cell culture, look to dissolved oxygen do2 for signs of bioeactor contamination. Mycoplasmas are not benign culture contaminants but have the ability to alter culture cell function, growth, metabolism, morphology, virus. No cell culture problem is as universal as that of culture loss due to contamination. Tissue culture waste disposal guide weill cornell medicine. It discusses how to reduce cell culture accidents by using good aseptic technique, the importance of freeze stocks, and how to appropriately monitor for contamination. Although cell culture contamination typically conjures thoughts of bacteria amongst cells and cloudy media, unwanted invaders in the culture flask can take many forms.
As mycoplasmal infection of cell cultures might often persist for long periods of time without apparent cell damage 4, it is important to use one or several efficient detection methods 1,3,5,6. The most common contaminants of cell cultures are mycoplasma, torula sp. The drug resistant cell line was actually identical to a ovarian cancer ovcar8 cell line in the panel. You can also try plating the culture medium onto some agar medium like ypd at each passage to check for yeast.
Cell culture supernatants, clinical specimens vaginal swabs, sputum, cryopreserved heart valve tissues, and reference strains were tested for mollicute contamination as well as to exclude cross. Cell culture contamination technical resources life. Request pdf on feb 1, 2004, simon p langdon and others published cell culture contamination. Cell culture basics handbook thermo fisher scientific. Contaminants of planttissue and cell cultures springerlink. The simple ways to prevent cell culture contamination. B oral mucosal epithelial cells derived from the patient after cell preparation. Cell culture contamination thermo fisher scientific us. Wear gloves but note that these are to protect you, and are not. Yeast contamination, when can i say ive defeat them. Understanding and managing cell culture contamination. Dec 26, 2012 for animal cell culture, look to dissolved oxygen do2 for signs of bioeactor contamination. Uncontaminated cell cultures uncontaminated cell cultures do not appear to present a significant hazard as even direct dermal inoculation may result in only local inflammation. Although it is nearly impossible to completely eliminate all contaminations, it is very important to minimize the frequency and.
Cell culture contamination stock photos download 53 royalty. Design and equipment for the cell culture laboratorycontents. Thus it is likely that mycoplasmas established themselves in cell culture from the very earliest attempts at in vitro cultivation and manipulation of cells as, by the time of the first published report, many workers checking their own cell lines as a consequence found they too had mycoplasma contamination. Animal cell culture 7 cell culture in media duration. Prevention and control of mycoplasma contamination in cell. Contamination in tissue culture covers the sources, prevention, detection, and elimination of contamination in tissue culture.
This webinar will introduce basic concepts of cell culture and provide tips on how to succeed in culturing mammalian cells. Crosscontamination of a cell culture with other cell types is a serious problem that has only recently been considered alarming. There are three main categories of cell culture contaminants. Aseptic technique is a combination of procedures designed to reduce the probability of infection. The most effective way to reduce the risk of biological contaminants is the proper use of aseptic. Both forms of contamination are extremely prevalent and cannot be underestimated. Hs324 disinfection of tissueculture waste guideline page 5 of 7 version.
The issue of cell line misidentification and cross contamination has been. All cell culture laboratories and cell culture workers have experi enced it. Jun 11, 20 cell culture is a fundamental technique for a variety of applications. At altogen labs, we provide services to rescue and decontaminate e. Work with cell cultures may also be covered by the requirements of the genetically modified organisms contained use regulations. Composed of 12 chapters, the book describes the frequency of occurrence of contamination and the many different effects of. Chemical contamination, often overlooked as a cause of cell growth problems, is caused by nonliving substances that produce unwanted effects on a culture system. In tissue culture medium, the embryoids formed from pollen grains is due to a organogenesis b cellular totipotency c double fertilization d test tube culture. However, 3070 days following cell culture contamination the increase in chromosomal aberrations was observed in the contaminated cell line. Virus contaminations of cell cultures a biotechnological view. Contamination of your cell culture can be the result of any of these conditions. Nov 18, 2015 animal cell culture 11 contamination by bacteria and yeast duration. Good luck, but be warned that this could be tricky. Cell cultures can become contaminated by the following means.
Thus it is likely that mycoplasmas established themselves in cell culture from the very earliest attempts at invitro cultivation and manipulation of cells as, by the time of the first published report, many workers checking their own cell lines as a consequence found they too had mycoplasma contamination. Nowadays, cell culture media are usually supplemented with the antibiotics, but first effect of antibiotics on cell cultured in vitro was established in the 1940s. When cultures become infected with microorganisms, or crosscontaminated by foreign cells, these cultures usually must be destroyed. Hs324 disinfection of tissueculture waste guideline page 1 of 7 version.
All materials that will come into direct contact with the culture must be sterile. At least 11 to 15% of cell cultures are currently infected by mycoplasmas. Contamination contamination is a staple problem in cell culture all cell culture laboratories and workers have experienced contamination affects both the use of cultures and research quality most apparent consequences of contamination. Pdf prevention and detection of mycoplasma contamination in. A novel bacterial contamination in cell culture manufacturing. Aug 20, 2014 good cell culture practices dictate that animal cells be grown without contamination by advantageous agents. All cell culture laboratories and cell culture workers have experienced it.
Understanding and managing cell culture contamination technical. With the number of researchers adding cell culture to their repertoire of techniques expanding daily, we believe there are many who can bene. Good cell culture practices dictate that animal cells be grown without contamination by advantageous agents. A method is described for eliminating fungal and other forms of contamination from valuable monolayer cell cultures. Contamination is a staple problem in cell culture all cell culture laboratories and workers have experienced contamination affects both the use of cultures and research quality most apparent consequences of contamination.
I just recently started cell culture and got my first cells in and already something seems fishy. For the cell culturist, two types of contamination require careful monitoring and constant vigilance. The most frequent causes of cell culture contamination are poor techniques and housekeeping and inadequate sterility testing of supplies and culture media. A population of animal cells will double once per day specific growth rate 0. Composed of 12 chapters, the book describes the frequency of occurrence of contamination and the many different effects of contamination on cultured cells. Culture contaminants may be biological or chemical, seen or unseen, destructive or seemingly benign, but in all cases they adversely affect both the use of your cell cultures and the quality. Doing in laboratory room bacterial contamination on agar media. Microbial contamination of cell cultures is a serious and relentless threat to your research. When they attempt to study the effects of pplo on hela cells. The relationship between culture maintenance conditions and the presence of mycoplasma were analyzed, considering the use of antibiotics in the culture media, fetal calf serum fcs quality, culture media processing, use of disponsable labware, type of laminar flow cabinet, quantity of operators, and cell culture system. Biological contamination is the dread of every per son working with cell culture.
Routine procedures used in this laboratory to control or prevent accidental contamina. Prepare the cell culture medium in biosafety cabinet a scientist looking cell morphology of cell culture. If the antibioticfree cultures remains sterile at 3 consecutive passages you should be fine. To view our online version and to request a copy of our sigmaecacc fundamental techniques in cell culture. Safety considerations in the cell culture laboratory.
Cell culture supernatants, clinical specimens vaginal swabs, sputum, cryopreserved heart valve tissues, and reference strains were tested for mollicute contamination as. Unknown fibrous contaminant in a human cell culture, visualized by scanning electron microscopy cell culture medium. Hela is a cervical adenocarcinoma cell derived from an africanamerican donor, henrietta lacks. Monitor rate of co2 use in incubators to determine when to change tanks. Contamination of cell cultures by bacteria including mycoplasmas, viruses, and fungi, or even crosscontamination by other cell lines, can result in a significant loss of resources for any research or pharmaceutical laboratory. Furthermore, coshh includes cell cultures in the definition of a biological agent. The third part of our lecture series on cell culture contamination focuses on everyday management of cell culture contamination. Segregate culture and test for mycoplasma infection. The failure was defined as potential contamination of cell culture production with microorganisms that have limited detectability with the current control systems l. Earle isolated mouse l fibroblasts which formed clones from single cells. In 2001 it was discovered that the breast cancer cell line mcf7 and its drug resistant daughter cell line mcf7 adrr adriamycin resistant were unrelated. Hopefully good technique, and not the use of antibiotics contribute to successful. An improved method of eliminating fungal contamination.
How to safeguard your cell culture from contamination contamination of your cell culture can be the result of any of these conditions. Cell culture contamination series corning life sciences. Primary cell types and stable cell lines are costly assets that are essential research reagents. Each possible source has a possible risk of transferring different types of contamination. How to safeguard your cell culture from contamination. Esco how to safeguard your cell culture from contamination. In addition to products, we provide a wealth of technical information, both online and in print, to help you reduce your risk of contamination. Cell culture contamination stock photos download 53. Animal and certainly plant cells grow more slowly than bacterial cells. Viral and chemical contaminants can also have considerable consequences for culture health, and ensuring that cell lines are not crosscontaminated is critical to reproducibility of results. Animal cell culture 11 contamination by bacteria and yeast duration. The cell line cross contamination and coculture is a major issue in animal cell culture that invalidates the research results, compromises the comparison of results in different laboratories and diminishes the use of animal cell culture for medical purpose and as a viable alternative and an effective tool in understanding the fundamental cell. The polio virus was cultured in simian and human kidney cells. Labs that have in place a good plan to reduce cell culture contaminations will usually have a much lower incidence of cell contamination than those labs that do not have such a plan.
Cell culture contamination technical resources life sciences. Maximum application of animal cell culture technology today is in the production of a insulin b interferons c edible proteins d vaccines. Cell culture contaminants can be divided into two main categories, chemical contaminants such as impurities in media, sera, and water, endotoxins, plasticizers, and detergents, and biological contaminants such as bacteria, molds, yeasts, viruses, mycoplasma, as well as cross contamination by other cell lines. Before conducting a clinical study, a researcher starts with primary steps like animal experiments mouse, nonhuman primates, etc. Virus contaminations of cell cultures a biotechnological. Mycoplasmapositive cell cultures are themselves the major source of. They both grow, but in the bhk21 flasks i see a lot of cells detaching and swimming in the medium. Hs324 disinfection of tissue culture waste guideline.
Can grow to high densities without any visible signs of contamination. The fta evaluated the following six categories of potential failure pathways. Secondary contamination may be present in aliquots prepared in the laboratory. When cultures become infected with microorganisms, or. Incubate this flask for at least one week to check for yeast growth then discard. Invasive mycoplasma, bacteria, and fungi can kill or drastically alter. Aliquot from each working cell bank wcb ampoule thaw preharvest cell culture fluid phccf optimized and implemented a commercial leptospiraspecific pcr assay to enhance detection sensitivity. Geyestablished a continuous cell line from a human cervical. Viral and chemical contaminants can also have considerable consequences for culture health, and ensuring that cell lines are not crosscontaminated is critical to. Cell culture contamination part 1 understanding cell.740 1038 58 580 357 71 1468 165 171 879 1403 958 29 492 1383 942 177 148 1474 1206 1532 1433 682 983 1036 661 269 229 236 1063 640 611 1354 440 590 577