Ann Dermatol.  2015 Jun;27(3):269-274. 10.5021/ad.2015.27.3.269.

Esterase Activity and Intracellular Localization in Reconstructed Human Epidermal Cultured Skin Models

• Tokudome Y
• Katayanagi M
• Hashimoto

• KMID: 2352497
• DOI: http://doi.org/10.5021/ad.2015.27.3.269

Abstract

BACKGROUND
Reconstructed human epidermal culture skin models have been developed for cosmetic and pharmaceutical research.
OBJECTIVE
This study evaluated the total and carboxyl esterase activities (i.e., K(m) and V(max), respectively) and localization in two reconstructed human epidermal culture skin models (LabCyte EPI-MODEL [Japan Tissue Engineering] and EpiDerm [MatTek/Kurabo]). The usefulness of the reconstruction cultured epidermis was also verified by comparison with human and rat epidermis.
METHODS
Homogenized epidermal samples were fractioned by centrifugation. p-nitrophenyl acetate and 4-methylumbelliferyl acetate were used as substrates of total esterase and carboxyl esterase, respectively.
RESULTS
Total and carboxyl esterase activities were present in the reconstructed human epidermal culture skin models and were localized in the cytosol. Moreover, the activities and localization were the same as those in human and rat epidermis.
CONCLUSION
LabCyte EPI-MODEL and EpiDerm are potentially useful for esterase activity prediction in human epidermis.

Keyword

Carboxyl esterase; Localization; Reconstructed human epidermal culture skin model; Skin esterase; Total esterase

MeSH Terms

Animals
Centrifugation
Cytosol
Epidermis
Humans
Rats
Skin*

Figure

• 

  Fig. 1 Effects of heat separation of rat epidermis and dermis on esterase activity. Left bar: intracellular localization of esterase activity in the whole epidermis without heat separation. Right bar: intracellular localization of esterase activity in the epidermis and dermis after heat separation. The esterase activities in epidermis and dermis are also shown. Data are mean±standard deviation (n=3).

• 

  Fig. 2 Total esterase activities of various skin samples. Mitochondria (M), light mitochondria (L), microsomal (P) and cytosolic (S) fractions. (A) Rat epidermis, (B) human epidermis, (C) LabCyte EPI-MODEL, (D) EpiDerm.

• 

  Fig. 3 Total esterase activities in the microsomal and cytosolic fractions of various skin samples. Data are mean±standard deviation (n=3).

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  Fig. 4 Carboxyl and aryl esterase activities in the microsomal and cytosolic fractions of various skin samples. (A) Carboxyl esterase, (B) aryl esterase. Data are mean±standard deviation (n=3). ND: not detected.

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