Human keratinocyte isolation and cell culture: a survey of current practices in the UK

A survey was conducted to establish current techniques for isolation and culture of human keratinocytes. A questionnaire was sent to all units thought to be involved in keratinocyte culture, a total of 34 individuals; 62 per cent of those surveyed responded to the questionnaires. The proportion of individuals using high-calcium medium to culture keratinocytes was 53 per cent, while 47 per cent used low-calcium serum-free medium. The majority of replies followed trends dependent on the culture method employed. Details of anatomical donor skin site, keratinocyte isolation and culture were compared. In particular, the problems associated with the use of commercially prepared low-calcium, serum-free medium were reported. The basic principles of keratinocyte culture reported in the literature were seen amongst all the replies received. It is interesting to note the variations in methods adopted as techniques are passed on and continuously modified to suit the requirements of the individual worker. This survey also highlights the difficulties that can occur when using mass-produced complex media.