Cell culture techniques【DOC精选】.docVIP

Cell culture techniques? /vaccines/en/poliolab/webhelp/ 4. Cell culture techniques? The quality of cell cultures used in the virological investigation of poliomyelitis is important for the standardization of poliovirus isolation and its characterization. This chapter provides some guidance on the correct handling of cell cultures used for virus diagnostic procedures. 4.1 Working in the cell culture laboratory Maintenance of trouble-free cell cultures depends on careful attention to culture conditions and passage procedures. It is also vital to pay strict attention to three characteristics that are fundamental to the quality of cell culture assays: purity, authenticity and stability. Purity: Contamination with microorganisms such as bacteria and fungi will normally kill the cells and put other cultures in the laboratory at risk. Mycoplasma contamination can have serious effects on a cell culture (see below) without inhibiting cell growth and, furthermore, the presence of such contamination will rarely be apparent even under microscopic observation. This is due to the extremely small size of mycoplasma organisms that can enable them to pass through sub-micron filters. As with bacteria and fungi, mycoplasma can spread readily to other cultures but are not susceptible to many of the antibiotics effective against bacterial contamination. While viral contamination typically produces a cytopathic effect in cell cultures, persistent non-cytopathic infections may arise that can influence virological investigations and may represent a hazard to laboratory workers (e.g. Epstein Barr Virus expressed by B95-8 and B95a cells). Screening for viral contamination can be extremely costly and time consuming. Routine checks for bacteria, fungi and mycoplasma, however, are relatively easy to establish and will provide confidence in the quality of cell culture results. Authenticity: Accidental switching of cell lines or cross-contamination between cultures has been identified in nu