HUMAN KERATINOCYTE CULTURE FROM THE PERITONSILLAR MUCOSA

Tonsillectomy tissue can be used as a routine source for cultures of o ropharyngeal keratinocytes. In so doing, a peritonsillar strip of unal tered mucosa was dissected in the upper submucosa. Subsequent trypsini zation yielded 7.0 +/- 3.4 x 10(6) keratinocytes per bilateral tonsill ectomy. Keratinocyte attachment and growth in primary culture were pro moted by sublethally irradiated 3T3 murine fibroblasts. Three subcultu res could be performed without a feeder layer and were characterized b y a population doubling time of 4.5 days during log growth phase. Elec trophoretic and immunoblot analysis of the third subculture revealed a strong expression of keratin pairs 5/14 and 6/16 as well as keratins 7 and 19, whereas keratins 8/18 were expressed less intensely. The low est intensity was found for keratin 13, which is known to be indicativ e of the differentiated mucosa. The culture technique thus provides an easily available in vitro model for morphological and functional stud ies on the epithelial compartment of human oropharyngeal mucosa.