Repeatable in-vitro mammalian cell culture requires pure and high quality culture media, clean and healthy cell lines, a precise and well maintained CO2 incubator, and good general aseptic culture technique. Major culture categories include:
- General small volume research
- Small volume research on structural cells requiring hypoxic growth conditions
- Medium-volume research on adherent cells
- High volume research and scale-up on adherent cells
- High volume research and scale-up on suspended cells
Only general government and best-practice guidelines have been applied to this type of research. The one exception is human In Vitro fertilization, which is regulated by government bodies such as the US FDA. Generally, incubators marketed for this purpose within the United States must be registered with the FDA as a Class II device.
To assure consistent and meaningful results, incubators used for these types of research need to feature tight parameter control, uniformity and stability over long periods of time. Depending on type of cell being cultured, parameters may include elevated CO2, suppressed O2, elevated temperature, and elevated humidity. Common issues to avoid include chamber contamination and loss of temperature control. The former is typically caused by contaminated cell lines or media, poor transfer technique, or insufficient incubator cleaning/decontamination/sterilization. The latter is a particular issue in non-refrigerated chambers being used with heat-generating cell agitation equipment (rollers, rockers, shakers, stirrers, etc) .
Other types of cell culture, including bacteriological and non-mammalian in-vitro, are often performed in similar chambers. CO2 is usually not required, but refrigeration may be for lower temperature ectothermic organisms. Humidification is usually not required for bacteria, given their short culture cycles.