caa a22 4500 606154620 CHVBK 20210128100559.0 cr unu---uuuuu 210128e20150901xx s 000 0 eng 10.1007/s10561-014-9485-2 doi (NATIONALLICENCE)springer-10.1007/s10561-014-9485-2 Production of an acellular matrix from amniotic membrane for the synthesis of a human skin equivalent [Elektronische Daten] [Anahí Sanluis-Verdes, Mª Yebra-Pimentel Vilar, Juan García-Barreiro, Marta García-Camba, Jacinto Ibáñez, Nieves Doménech, Mª Rendal-Vázquez] 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 30min 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. Springer Science+Business Media Dordrecht, 2015 Skin equivalent nationallicence Human amniotic membrane nationallicence Keratinocytes nationallicence Biomaterial nationallicence Trypsin nationallicence Sanluis-Verdes Anahí 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 aut Yebra-Pimentel Vilar Mª 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 aut García-Barreiro Juan 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 aut García-Camba Marta 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 aut Ibáñez Jacinto 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 aut Doménech Nieves 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 aut Rendal-Vázquez Mª 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 aut Cell and Tissue Banking Springer Netherlands 16/3(2015-09-01), 411-423 1389-9333 16:3<411 2015 16 10561 https://doi.org/10.1007/s10561-014-9485-2 text/html Onlinezugriff via DOI BK010053 XK010053 XK010000 Metadata rights reserved Springer special CC-BY-NC licence nationallicence 1 research-article jats NATIONALLICENCE NATIONALLICENCE NL-springer NATIONALLICENCE 856 40 https://doi.org/10.1007/s10561-014-9485-2 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Sanluis-Verdes Anahí 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 aut NATIONALLICENCE 700 1- Yebra-Pimentel Vilar Mª 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 aut NATIONALLICENCE 700 1- García-Barreiro Juan 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 aut NATIONALLICENCE 700 1- García-Camba Marta 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 aut NATIONALLICENCE 700 1- Ibáñez Jacinto 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 aut NATIONALLICENCE 700 1- Doménech Nieves 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 aut NATIONALLICENCE 700 1- Rendal-Vázquez Mª 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 aut NATIONALLICENCE 773 0- Cell and Tissue Banking Springer Netherlands 16/3(2015-09-01), 411-423 1389-9333 16:3<411 2015 16 10561