Human keratinocyte cultures as models of cutaneous esterase activity

Toxicology in Vitro : an International Journal Published in Association with BIBRA
C L Barker, R Clothier

Abstract

A reproducible, quantifiable assay has been developed for the measurement of esterase activity in human keratinocyte cultures, using the model substrate 4-methyl umbelliferyl heptanoate (MUH) which is hydrolysed to the fluorescent metabolite 4-methyl umbelliferone (MU). Activity was assessed in two human keratinocyte cell lines, NCTC 2544 and SVK-14, and in freshly isolated human breast keratinocytes from primary culture to passage 3. V(max) values for MUH hydrolysing activity in the two cell lines showed that the less differentiated cell line NCTC 2544 (V(max) = 23.00 +/- 2.84) expresses a much higher activity than SVK-14s (V(max) = 13.28 +/- 1.42) which are more differentiated and able to form a cornified envelope. Activity in the freshly isolated human breast keratinocytes decreased with time in culture in all three donors tested, which is also likely to relate to the extent of cell differentiation. In human skin, xenobiotic esters penetrating the stratum corneum may be exposed to changing levels of hydrolysing esterases as they are absorbed across the epidermal cell layers. The assay for MUH hydrolysis will be a useful tool for the study of esterase activity in populations of human keratinocytes in vitro.

References

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Citations

Sep 22, 2001·Toxicology in Vitro : an International Journal Published in Association with BIBRA·A C Gray, R H Clothier
Sep 22, 2001·Toxicology in Vitro : an International Journal Published in Association with BIBRA·L ReidN Khammo
Aug 1, 1999·Toxicology in Vitro : an International Journal Published in Association with BIBRA·A C GrayR H Clothier
Apr 5, 2012·Journal of Immunotoxicology·Christine JäckhRobert Landsiedel
Dec 17, 2014·European Journal of Pharmaceutics and Biopharmaceutics : Official Journal of Arbeitsgemeinschaft Für Pharmazeutische Verfahrenstechnik E.V·Joseph A LaszloDavid L Compton
Jun 17, 2006·Skin Pharmacology and Physiology·K AbdulmajedC M Heard
Feb 1, 2011·Alternatives to Laboratory Animals : ATLA·Richard Clothier
Aug 1, 2007·Drug Metabolism and Disposition : the Biological Fate of Chemicals·Christopher JewellFaith M Williams
Jul 30, 2019·Skin Pharmacology and Physiology·Sung Min Pyo, Howard I Maibach

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