Model of human epidermis reconstructed in vitro with keratinocytes and melanocytes on dead de-epidermized human dermis

Authors

  • Jussara Rehder Faculdade de Ciências Médicas, Universidade Estadual Campinas
  • Luís Ricardo Martinhão Souto Faculdade de Ciências Médicas, Universidade Estadual Campinas
  • Cláudia Maria Bernardino Magro Issa Faculdade de Ciências Médicas, Universidade Estadual Campinas
  • Maria Beatriz Puzzi Faculdade de Ciências Médicas, Universidade Estadual Campinas

Keywords:

Epidermis, Melanocyte, Culture, Keratinocyte

Abstract

CONTEXT: Recent progress in the field of epithelial culture techniques has allowed the development of culture systems in which the reconstructed epidermis presents characteristics of morphological differentiation similar to those seen in vivo. Human epidermis reconstructed in vitro may be used as the best alternative for the in vitro testing of the toxicology and efficiency of products for topical use, as well as in the treatment of skin burns and chronic skin ulcers. OBJECTIVE: To demonstrate a method for obtaining human epidermis reconstructed in vitro, using keratinocytes and melanocytes cultivated on dead de-epidermized human dermis. TYPE OF STUDY: Experimental/laboratory. SETTING: Skin Cell Culture Laboratory of the Faculdade de Ciências Médicas, Universidade Estadual de Campinas, Campinas, São Paulo, Brazil. PROCEDURE: Human keratinocytes and melanocytes cultured in vitro were grown on a biological matrix (dead de-epidermized human dermis) and the system was kept at an air-liquid interface, in a suitable culturing medium, until a stratified human epidermis was formed, maintaining the histological characteristics of the epidermis in vivo. RESULTS: It was histologically demonstrated that it is possible to reproduce a differentiated epidermis through keratinocytes and melanocytes cultured on dead de-epidermized human dermis, thus obtaining a correctly positioned human epidermis reconstructed in vitro with functional keratinocytes and melanocytes that is similar to in vivo epidermis. CONCLUSIONS: It is possible to obtain a completely differentiated human epidermis reconstructed in vitro from keratinocyte and melanocyte cultures on a dead de-epidermized human dermis.

Downloads

Download data is not yet available.

Author Biographies

Jussara Rehder, Faculdade de Ciências Médicas, Universidade Estadual Campinas

MD. Chief biologist of the Laboratory of Molecular Biology and Skin Cell Culture Laboratory, Universidade Estadual de Campinas, Campinas, São Paulo, Brazil.

Luís Ricardo Martinhão Souto, Faculdade de Ciências Médicas, Universidade Estadual Campinas

MD. Plastic surgeon and MSc student of Medical Sciences at Faculdade de Ciências Médicas, Universidade Estadual de Campinas, Campinas, São Paulo, Brazil.

Cláudia Maria Bernardino Magro Issa, Faculdade de Ciências Médicas, Universidade Estadual Campinas

MD. Dermatologist and PhD student of Internal Medicine at Faculdade de Ciências Médicas, Universidade Estadual de Campinas, Campinas, São Paulo, Brazil.

Maria Beatriz Puzzi, Faculdade de Ciências Médicas, Universidade Estadual Campinas

MD, PhD. Professor of the Discipline of Dermatology, Department of Internal Medicine, and Head of the Skin Cell Culture Laboratory at Faculdade de Ciências Médicas, Universidade Estadual de Campinas, Campinas, São Paulo, Brazil.

References

Boranic M, Jakic-Razumovic J, Stanovic S, Kljenak A, Fattorini I. Kultura koznih stanica: primjena u plasticnoj kirurgiji i laboratorijskom istrazivanju. [Skin cell culture: utilization in plastic surgery and laboratory studies]. Lijec Vjesn 1999;121(4- 5):137-43.

Watt FM. The epidermal keratinocyte. Bioessays 1988; 8(5):163-7.

Taube MBP, Taieb A. Metabolismo lipídico na cultura de queratinócitos. [Lipid metabolism in cultured keratinocytes]. An Bras Dermatol 2000;75(1):75-84.

Huang Y, Ren L, Qin Y. Observation of cicatricial fibroblasts in culture and its biological properties. Zhongguo Xiu Fu Chong Jian Wai Ke Za Zhi 1998;12(6):332-5.

Valyi-Nagy IT, Murphy GF, Mancianti ML, Whitaker D, Herlyn M. Phenotypes and interactions of human melanocytes and keratinocytes in an epidermal reconstruction model. Lab In- vest 1990;62(3):314-24.

Bernerd F, Asselineau D. Successive alteration and recovery of epidermal differentiation and morphogenesis after specific UVB- damages in skin reconstructed in vitro. Dev Biol 1997;183(2):123-38.

Régnier M, Patwardhan A, Scheynius A, et al. Reconstructed human epidermis composed of keratinocytes, melanocytes and Langerhans cells. Med Biol Eng Comput 1998;36(6):821-4.

Chistolini P, De Angelis G, De Luca M, Pellegrini G, Ruspantini I. Analysis of the mechanical properties of in vitro reconstructed epidermis: preliminary results. Med Biol Eng Comput 1999;37(5):670-2.

Carsin H, Ainaud P, Le Bever H, et al. Cultured epithelial autografts in extensive burn coverage of severely traumatized patients: a five year single-center experience with 30 patients. Burns 2000;26(4):379-87.

Régnier M, Caron D, Reichert U, Schaefer H. Reconstructed human epidermis: a model to study in vitro the barrier func- tion of the skin. Skin Pharmacol 1992;5(1):49-56.

Régnier M, Asselineau D, Lenoir MC. Human epidermis re- constructed on dermal substrates in vitro: an alternative to ani- mals in skin pharmacology. Skin Pharmacol 1990;3(2):70-85.

Pruniéras M, Régnier M, Woodley D. Methods for cultivation of keratinocytes with an air-liquid interface. J Invest Dermatol 1983;81(1 Suppl):28s-33s.

Ponec M, Gibbs S, Pilgram G, et al. Barrier function in recon- structed epidermis and its resemblance to native human skin. Skin Pharmacol Appl Skin Physiol 2001;14(Suppl 1):63-71.

van Dorp AG, Verhoeven MC, Nat-Van Der Meij TH, Koerten HK, Ponec M. A modified culture system for epidermal cells for grafting purposes: an in vitro and in vivo study. Wound Repair Regen 1999;7(4):214-25.

Bessou S, Surlève-Bazeille JE, Sorbier E, Taieb A. Ex vivo re- construction of the epidermis with melanocytes and the influ- ence of UVB. Pigment Cell Res 1995;8(5):241-9.

Pruniéras M, Régnier M, Schlotterer M. Nouveau procédé de culture des cellules épidermiques humaines sur derme homo- logue ou hétérologue: préparation de greffons recombinés. [New procedure for culturing human epidermal cells on allogenic or xenogenic skin: preparation of recombined grafts]. Ann Chir Plast 1979;24(4):357-62.

Régnier M, Pruniéras M, Woodley D. Growth and differentia- tion of adult human epidermal cells on dermal substrates. Front Matrix Biol 1981;9:4-35.

Terskikh VV, Vasiliev AV. Cultivation and transplantation of epidermal keratinocytes. Int Rev Cytol 1999;188:41-72.

Ponec M, Kempenaar J, Weerheim A, de Lannoy L, Kalkman I, Jansen H. Triglyceride metabolism in human keratinocytes cul- tured at the air-liquid interface. Arch Dermatol Res 1995;287(8):723-30.

Downloads

Published

2004-01-01

How to Cite

1.
Rehder J, Souto LRM, Issa CMBM, Puzzi MB. Model of human epidermis reconstructed in vitro with keratinocytes and melanocytes on dead de-epidermized human dermis. Sao Paulo Med J [Internet]. 2004 Jan. 1 [cited 2025 Oct. 16];122(1):22-5. Available from: https://periodicosapm.emnuvens.com.br/spmj/article/view/2483

Issue

Section

Original Article