Calcium: a crucial consideration in serum‐free keratinocyte culture

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Abstract
This investigation was conducted when previously rcpealable experimental data became impossible to reproduce when using keratino‐cytes cultured in serum‐free medium. Differences in calcium molarity between batches of medium were identified as a source of variation in cultured keratinocyte populations. The susceptibility of cultured keralinocytes to even small alterations in calcium molarily has been demonstrated. 2 regular medium batches were compared with a special preparation of medium, devoid of calcium chloride then supplemented with a known concentration of calcium ions. Culture progress in each medium was assessed by: morphological observation, % cells expressing involucrin and proliferating cell nuclear antigen, cell attachment, growth rate and colony forming efficiency. In order to control the phenotype of cultured kera‐tinocytes, in a rcpoducible system, it is recommended that serum‐free keratinocyte medium is purchased with the omission of calcium chloride. Supplementation of this medium may then be made by the investigator to suit individual culture requirements.