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Production of an acellular matrix from amniotic membrane for the synthesis of a human skin equivalent

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Abstract

Human amniotic membrane (HAM) has useful properties as a dermal matrix substitute. The objective of our work was to obtain, using different enzymatic or chemical treatments to eliminate cells, a scaffold of acellular HAM for later use as a support for the development of a skin equivalent. The HAM was separated from the chorion, incubated and cryopreserved. The membrane underwent different enzymatic and chemical treatments to eliminate the cells. Fibroblasts and keratinocytes were separately obtained from skin biopsies of patients following a sequential double digestion with first collagenase and then trypsin–EDTA (T/E). A skin equivalent was then constructed by seeding keratinocytes on the epithelial side and fibroblasts on the chorionic side of the decellularizated HAM. Histological, immunohistochemical, inmunofluorescent and molecular biology studies were performed. Treatment with 1 % T/E at 37 °C for 30 min totally removed epithelial and mesenchymal cells. The HAM thus treated proved to be a good matrix to support adherence of cells and allowed the achievement of an integral and intact scaffold for development of a skin equivalent, which could be useful as a skin substitute for clinical use.

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Acknowledgments

This work was supported by a grant from “Consellería de Innovación e Industria Dirección Xeral de I+D+I (No. 08CSA065916PR)”. A. Verdes-Sanluis was supported by a grant from “Centro de Investigación Biomédica en Red en Bioingeniería, Biomateriales y Nanomedicina” (CIBER-BBN-CB06/01/0040).

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Authors and Affiliations

  1. Unidad de Criobiología-Banco de Tejidos, Instituto de Investigación Biomédica de A Coruña (INIBIC), Complexo Hospitalario Universitario de A Coruña (CHUAC), Sergas, As Xubias, s/n, 15006, A Coruña, Spain

    Anahí Sanluis-Verdes, Jacinto Sánchez Ibáñez & Mª Esther Rendal-Vázquez

  2. Servicio de Anatomía Patológica, Instituto de Investigación Biomédica de A Coruña (INIBIC), Complexo Hospitalario Universitario de A Coruña (CHUAC), Sergas, As Xubias, s/n, 15006, A Coruña, Spain

    Mª Teresa Yebra-Pimentel Vilar

  3. Unidad de Quemados, Instituto de Investigación Biomédica de A Coruña (INIBIC), Complexo Hospitalario Universitario de A Coruña (CHUAC), Sergas, As Xubias, s/n, 15006, A Coruña, Spain

    Juan Javier García-Barreiro

  4. Biobanco A Coruña, Instituto de Investigación Biomédica de A Coruña (INIBIC), Complexo Hospitalario Universitario de A Coruña (CHUAC), Sergas, As Xubias, s/n, 15006, A Coruña, Spain

    Marta García-Camba & Nieves Doménech

Authors
  1. Anahí Sanluis-Verdes
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  2. Mª Teresa Yebra-Pimentel Vilar
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  3. Juan Javier García-Barreiro
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  4. Marta García-Camba
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  5. Jacinto Sánchez Ibáñez
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  6. Nieves Doménech
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  7. Mª Esther Rendal-Vázquez
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Corresponding authors

Correspondence to Nieves Doménech or Mª Esther Rendal-Vázquez.

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Sanluis-Verdes, A., Yebra-Pimentel Vilar, M.T., García-Barreiro, J.J. et al. Production of an acellular matrix from amniotic membrane for the synthesis of a human skin equivalent. Cell Tissue Bank 16, 411–423 (2015). https://doi.org/10.1007/s10561-014-9485-2

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  • DOI: https://doi.org/10.1007/s10561-014-9485-2

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