A sterile, liquid medium prepared without calcium chloride for the long-term, serum-free culture of human epidermal keratinocytes and human corneal epithelial cells. This basal medium requires the addition of calcium plus an appropriate supplement prior to use. Calcium chloride is provided as a separate component with each bottle of medium. EpiLife greatly extends the in vitro lifespan of human epidermal keratinocytes compared to other serum-free media systems. This basal media is animal origin free and contains essential and non-essential amino acids, vitamins, other organic compounds, trace minerals, and inorganic salts. It does not contain antibiotics, antimycotics, hormones, growth factors, or proteins. EpiLife® medium is HEPES and bicarbonate buffered, and is designed for use in an incubator with an atmosphere of 5% CO2 and 95% air.