Novel Decellularization Method for Tissue Slices
Decellularization procedures have been developed and optimized for the entire organ or tissue blocks, by either perfusion of decellularizing agents through the tissue’s vasculature or submerging large sections in decellularizing solutions. However, some research aims require the analysis of native a...
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2022-03-01
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| Series: | Frontiers in Bioengineering and Biotechnology |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fbioe.2022.832178/full |
| _version_ | 1819294674953699328 |
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| author | Maria Narciso Maria Narciso Anna Ulldemolins Constança Júnior Constança Júnior Jorge Otero Jorge Otero Jorge Otero Daniel Navajas Daniel Navajas Daniel Navajas Ramon Farré Ramon Farré Ramon Farré Núria Gavara Núria Gavara Isaac Almendros Isaac Almendros Isaac Almendros |
| author_facet | Maria Narciso Maria Narciso Anna Ulldemolins Constança Júnior Constança Júnior Jorge Otero Jorge Otero Jorge Otero Daniel Navajas Daniel Navajas Daniel Navajas Ramon Farré Ramon Farré Ramon Farré Núria Gavara Núria Gavara Isaac Almendros Isaac Almendros Isaac Almendros |
| author_sort | Maria Narciso |
| collection | DOAJ |
| description | Decellularization procedures have been developed and optimized for the entire organ or tissue blocks, by either perfusion of decellularizing agents through the tissue’s vasculature or submerging large sections in decellularizing solutions. However, some research aims require the analysis of native as well as decellularized tissue slices side by side, but an optimal protocol has not yet been established to address this need. Thus, the main goal of this work was to develop a fast and efficient decellularization method for tissue slices—with an emphasis on lung—while attached to a glass slide. To this end, different decellularizing agents were compared for their effectiveness in cellular removal while preserving the extracellular matrix. The intensity of DNA staining was taken as an indicator of remaining cells and compared to untreated sections. The presence of collagen, elastin and laminin were quantified using immunostaining and signal quantification. Scaffolds resulting from the optimized protocol were mechanically characterized using atomic force microscopy. Lung scaffolds were recellularized with mesenchymal stromal cells to assess their biocompatibility. Some decellularization agents (CHAPS, triton, and ammonia hydroxide) did not achieve sufficient cell removal. Sodium dodecyl sulfate (SDS) was effective in cell removal (1% remaining DNA signal), but its sharp reduction of elastin signal (only 6% remained) plus lower attachment ratio (32%) singled out sodium deoxycholate (SD) as the optimal treatment for this application (6.5% remaining DNA signal), due to its higher elastin retention (34%) and higher attachment ratio (60%). Laminin and collagen were fully preserved in all treatments. The SD decellularization protocol was also successful for porcine and murine (mice and rat) lungs as well as for other tissues such as the heart, kidney, and bladder. No significant mechanical differences were found before and after sample decellularization. The resulting acellular lung scaffolds were shown to be biocompatible (98% cell survival after 72 h of culture). This novel method to decellularize tissue slices opens up new methodological possibilities to better understand the role of the extracellular matrix in the context of several diseases as well as tissue engineering research and can be easily adapted for scarce samples like clinical biopsies. |
| first_indexed | 2024-12-24T04:30:05Z |
| format | Article |
| id | doaj.art-cea8a612d7cb435493e8d3f0c9fe10ac |
| institution | Directory Open Access Journal |
| issn | 2296-4185 |
| language | English |
| last_indexed | 2024-12-24T04:30:05Z |
| publishDate | 2022-03-01 |
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| record_format | Article |
| series | Frontiers in Bioengineering and Biotechnology |
| spelling | doaj.art-cea8a612d7cb435493e8d3f0c9fe10ac2022-12-21T17:15:28ZengFrontiers Media S.A.Frontiers in Bioengineering and Biotechnology2296-41852022-03-011010.3389/fbioe.2022.832178832178Novel Decellularization Method for Tissue SlicesMaria Narciso0Maria Narciso1Anna Ulldemolins2Constança Júnior3Constança Júnior4Jorge Otero5Jorge Otero6Jorge Otero7Daniel Navajas8Daniel Navajas9Daniel Navajas10Ramon Farré11Ramon Farré12Ramon Farré13Núria Gavara14Núria Gavara15Isaac Almendros16Isaac Almendros17Isaac Almendros18Unitat de Biofísica i Bioenginyeria, Facultat de Medicina i Ciències de la Salut, Universitat de Barcelona, Barcelona, SpainThe Institute for Bioengineering of Catalonia (IBEC), The Barcelona Institute of Science and Technology, Barcelona, SpainUnitat de Biofísica i Bioenginyeria, Facultat de Medicina i Ciències de la Salut, Universitat de Barcelona, Barcelona, SpainUnitat de Biofísica i Bioenginyeria, Facultat de Medicina i Ciències de la Salut, Universitat de Barcelona, Barcelona, SpainThe Institute for Bioengineering of Catalonia (IBEC), The Barcelona Institute of Science and Technology, Barcelona, SpainUnitat de Biofísica i Bioenginyeria, Facultat de Medicina i Ciències de la Salut, Universitat de Barcelona, Barcelona, SpainThe Institute for Bioengineering of Catalonia (IBEC), The Barcelona Institute of Science and Technology, Barcelona, SpainCIBER de Enfermedades Respiratorias, Madrid, SpainUnitat de Biofísica i Bioenginyeria, Facultat de Medicina i Ciències de la Salut, Universitat de Barcelona, Barcelona, SpainThe Institute for Bioengineering of Catalonia (IBEC), The Barcelona Institute of Science and Technology, Barcelona, SpainCIBER de Enfermedades Respiratorias, Madrid, SpainUnitat de Biofísica i Bioenginyeria, Facultat de Medicina i Ciències de la Salut, Universitat de Barcelona, Barcelona, SpainCIBER de Enfermedades Respiratorias, Madrid, SpainInstitut d’Investigacions Biomèdiques August Pi i Sunyer, Barcelona, SpainUnitat de Biofísica i Bioenginyeria, Facultat de Medicina i Ciències de la Salut, Universitat de Barcelona, Barcelona, SpainThe Institute for Bioengineering of Catalonia (IBEC), The Barcelona Institute of Science and Technology, Barcelona, SpainUnitat de Biofísica i Bioenginyeria, Facultat de Medicina i Ciències de la Salut, Universitat de Barcelona, Barcelona, SpainCIBER de Enfermedades Respiratorias, Madrid, SpainInstitut d’Investigacions Biomèdiques August Pi i Sunyer, Barcelona, SpainDecellularization procedures have been developed and optimized for the entire organ or tissue blocks, by either perfusion of decellularizing agents through the tissue’s vasculature or submerging large sections in decellularizing solutions. However, some research aims require the analysis of native as well as decellularized tissue slices side by side, but an optimal protocol has not yet been established to address this need. Thus, the main goal of this work was to develop a fast and efficient decellularization method for tissue slices—with an emphasis on lung—while attached to a glass slide. To this end, different decellularizing agents were compared for their effectiveness in cellular removal while preserving the extracellular matrix. The intensity of DNA staining was taken as an indicator of remaining cells and compared to untreated sections. The presence of collagen, elastin and laminin were quantified using immunostaining and signal quantification. Scaffolds resulting from the optimized protocol were mechanically characterized using atomic force microscopy. Lung scaffolds were recellularized with mesenchymal stromal cells to assess their biocompatibility. Some decellularization agents (CHAPS, triton, and ammonia hydroxide) did not achieve sufficient cell removal. Sodium dodecyl sulfate (SDS) was effective in cell removal (1% remaining DNA signal), but its sharp reduction of elastin signal (only 6% remained) plus lower attachment ratio (32%) singled out sodium deoxycholate (SD) as the optimal treatment for this application (6.5% remaining DNA signal), due to its higher elastin retention (34%) and higher attachment ratio (60%). Laminin and collagen were fully preserved in all treatments. The SD decellularization protocol was also successful for porcine and murine (mice and rat) lungs as well as for other tissues such as the heart, kidney, and bladder. No significant mechanical differences were found before and after sample decellularization. The resulting acellular lung scaffolds were shown to be biocompatible (98% cell survival after 72 h of culture). This novel method to decellularize tissue slices opens up new methodological possibilities to better understand the role of the extracellular matrix in the context of several diseases as well as tissue engineering research and can be easily adapted for scarce samples like clinical biopsies.https://www.frontiersin.org/articles/10.3389/fbioe.2022.832178/fulldecellularizationbioscaffold recellularizationbiocompatibilityextracellular matrixtissue sliceslung |
| spellingShingle | Maria Narciso Maria Narciso Anna Ulldemolins Constança Júnior Constança Júnior Jorge Otero Jorge Otero Jorge Otero Daniel Navajas Daniel Navajas Daniel Navajas Ramon Farré Ramon Farré Ramon Farré Núria Gavara Núria Gavara Isaac Almendros Isaac Almendros Isaac Almendros Novel Decellularization Method for Tissue Slices Frontiers in Bioengineering and Biotechnology decellularization bioscaffold recellularization biocompatibility extracellular matrix tissue slices lung |
| title | Novel Decellularization Method for Tissue Slices |
| title_full | Novel Decellularization Method for Tissue Slices |
| title_fullStr | Novel Decellularization Method for Tissue Slices |
| title_full_unstemmed | Novel Decellularization Method for Tissue Slices |
| title_short | Novel Decellularization Method for Tissue Slices |
| title_sort | novel decellularization method for tissue slices |
| topic | decellularization bioscaffold recellularization biocompatibility extracellular matrix tissue slices lung |
| url | https://www.frontiersin.org/articles/10.3389/fbioe.2022.832178/full |
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