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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...

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Autores principales: Stanton, Alice E., Tong, Xinming, Yang, Fan
Formato: Online Artículo Texto
Lenguaje:English
Publicado: AIP Publishing LLC 2019
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.
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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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