Home banner
Divider
A-Z Index

Quick way to the find the information that you need...

More button
Register with FRAME

Although you do not need to register, any information you provide will be confidential and used only by FRAME to improve the website

Register button
Account Login
Forgot password?

ATLA - ISI
The Journal

 

Alternatives to Laboratory Animals - ATLA

Download latest issue button Download back issues button Subscribe to ATLA
Contact Us

Tel icon

Tel: +44 (0)115 9584740


Tel icon

Fax: +44 (0)115 9503570

Make an Enquiry

Human keratinocyte cultures as models of cutaneous esterase activity.


Barker, C.L. and Clothier, R.H.

Toxicology in Vitro, 11(5), 637-640 (1997).

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. Vmax values for MUH hydrolysing activity in the two cell lines showed that the less differentiated cell line NCTC 2544 (Vmax = 23.00 ± 2.84) expresses a much higher activity than SVK-14s (Vmax = 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.