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Varying solvent type modulates collagen coating and stem cell mechanotransduction on hydrogel substrates
Type I collagen is the most abundant extracellular matrix protein in the human body and is commonly used as a biochemical ligand for hydrogel substrates to support cell adhesion in mechanotransduction studies. Previous protocols for conjugating collagen I have used different solvents; yet, how varyi...
Autores principales: | , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
AIP Publishing LLC
2019
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6768796/ https://www.ncbi.nlm.nih.gov/pubmed/31592041 http://dx.doi.org/10.1063/1.5111762 |
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author | Stanton, Alice E. Tong, Xinming Yang, Fan |
author_facet | Stanton, Alice E. Tong, Xinming Yang, Fan |
author_sort | Stanton, Alice E. |
collection | PubMed |
description | Type I collagen is the most abundant extracellular matrix protein in the human body and is commonly used as a biochemical ligand for hydrogel substrates to support cell adhesion in mechanotransduction studies. Previous protocols for conjugating collagen I have used different solvents; yet, how varying solvent pH and composition impacts the efficiency and distribution of these collagen I coatings remains unknown. Here, we examine the effect of varying solvent pH and type on the efficiency and distribution of collagen I coatings on polyacrylamide hydrogels. We further evaluate the effects of varying solvent on mechanotransduction of human mesenchymal stem cells (MSCs) by characterizing cell spreading and localization of Yes-Associated Protein (YAP), a key transcriptional regulator of mechanotransduction. Increasing solvent pH to 5.2 and above increased the heterogeneity of coating with collagen bundle formation. Collagen I coating highly depends on the solvent type, with acetic acid leading to the highest conjugation efficiency and most homogeneous coating. Compared to HEPES or phosphate-buffered saline buffer, acetic acid-dissolved collagen I coatings substantially enhance MSC adhesion and spreading on both glass and polyacrylamide hydrogel substrates. When acetic acid was used for collagen coatings, even the low collagen concentration (1 μg/ml) induced robust MSC spreading and nuclear YAP localization on both soft (3 kPa) and stiff (38 kPa) substrates. Depending on the solvent type, stiffness-dependent nuclear YAP translocation occurs at a different collagen concentration. Together, the results from this study validate the solvent type as an important parameter to consider when using collagen I as the biochemical ligand to support cell adhesion. |
format | Online Article Text |
id | pubmed-6768796 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2019 |
publisher | AIP Publishing LLC |
record_format | MEDLINE/PubMed |
spelling | pubmed-67687962019-10-07 Varying solvent type modulates collagen coating and stem cell mechanotransduction on hydrogel substrates Stanton, Alice E. Tong, Xinming Yang, Fan APL Bioeng Articles Type I collagen is the most abundant extracellular matrix protein in the human body and is commonly used as a biochemical ligand for hydrogel substrates to support cell adhesion in mechanotransduction studies. Previous protocols for conjugating collagen I have used different solvents; yet, how varying solvent pH and composition impacts the efficiency and distribution of these collagen I coatings remains unknown. Here, we examine the effect of varying solvent pH and type on the efficiency and distribution of collagen I coatings on polyacrylamide hydrogels. We further evaluate the effects of varying solvent on mechanotransduction of human mesenchymal stem cells (MSCs) by characterizing cell spreading and localization of Yes-Associated Protein (YAP), a key transcriptional regulator of mechanotransduction. Increasing solvent pH to 5.2 and above increased the heterogeneity of coating with collagen bundle formation. Collagen I coating highly depends on the solvent type, with acetic acid leading to the highest conjugation efficiency and most homogeneous coating. Compared to HEPES or phosphate-buffered saline buffer, acetic acid-dissolved collagen I coatings substantially enhance MSC adhesion and spreading on both glass and polyacrylamide hydrogel substrates. When acetic acid was used for collagen coatings, even the low collagen concentration (1 μg/ml) induced robust MSC spreading and nuclear YAP localization on both soft (3 kPa) and stiff (38 kPa) substrates. Depending on the solvent type, stiffness-dependent nuclear YAP translocation occurs at a different collagen concentration. Together, the results from this study validate the solvent type as an important parameter to consider when using collagen I as the biochemical ligand to support cell adhesion. AIP Publishing LLC 2019-09-30 /pmc/articles/PMC6768796/ /pubmed/31592041 http://dx.doi.org/10.1063/1.5111762 Text en © Author(s). 2473-2877/2019/3(3)/036108/9 All article content, except where otherwise noted, is licensed under a Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Articles Stanton, Alice E. Tong, Xinming Yang, Fan Varying solvent type modulates collagen coating and stem cell mechanotransduction on hydrogel substrates |
title | Varying solvent type modulates collagen coating and stem cell mechanotransduction on hydrogel substrates |
title_full | Varying solvent type modulates collagen coating and stem cell mechanotransduction on hydrogel substrates |
title_fullStr | Varying solvent type modulates collagen coating and stem cell mechanotransduction on hydrogel substrates |
title_full_unstemmed | Varying solvent type modulates collagen coating and stem cell mechanotransduction on hydrogel substrates |
title_short | Varying solvent type modulates collagen coating and stem cell mechanotransduction on hydrogel substrates |
title_sort | varying solvent type modulates collagen coating and stem cell mechanotransduction on hydrogel substrates |
topic | Articles |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6768796/ https://www.ncbi.nlm.nih.gov/pubmed/31592041 http://dx.doi.org/10.1063/1.5111762 |
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