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Because they are derived directly from living tissue, primary cells maintain physiological relevance and thus find increasing use in life science research and pharmaceutical drug discovery. Compared to immortalized cell lines, however, primary cells can be finicky and present challenges during culture unless proper care is taken. Cell Applications recommends the user follow the steps outlined in these video segments to achieve optimal results with their primary cells. Prior to working with the cells, review the cell-specific steps and instructions provided by CAI. Adhere to standard, accepted guidelines for sterile technique, laboratory safety and equipment, such as appropriate eye and skin protection when moving the vials to and from liquid nitrogen storage. Work with the cells in a Class II Biological Safety Cabinet and culture them in a humidified incubator set to 37°C and 5% CO2. Turn on the water bath and set its temperature to 37°C at least one hour in advance, and check the temperature with a thermometer prior to thawing the cells. Ensure that you have the optimized, cell-specific growth media. Also, confirm that your cell culture flasks are suitable for primary cell culture. To reduce the chances for contamination, use 70% isopropyl alcohol to wipe down working surfaces, media bottles and thawed cyrovials.