Engineering Hydrogels with Affinity-Bound Laminin as 3D Neural Stem Cell Culture Systems
Autor(a) principal: | |
---|---|
Data de Publicação: | 2019 |
Outros Autores: | , , , , , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
Texto Completo: | https://hdl.handle.net/10216/122944 |
Resumo: | Laminin incorporation into biological or synthetic hydrogels has been explored to recapitulate the dynamic nature and biological complexity of neural stem cell (NSC) niches. However, the strategies currently explored for laminin immobilization within three-dimensional (3D) matrices do not address a critical aspect influencing cell-matrix interactions, which is the control over laminin conformation and orientation upon immobilization. This is a key feature for the preservation of the protein bioactivity. In this work, we explored an affinity-based approach to mediate the site-selective immobilization of laminin into a degradable synthetic hydrogel. Specifically, a four-arm maleimide terminated poly(ethylene glycol) (PEG-4MAL) macromer was functionalized with a mono-PEGylated recombinant human N-terminal agrin (NtA) domain, to promote high affinity binding of laminin. Different NtA concentrations (10, 50 and 100 M) were used to investigate the impact of NtA density on laminin incorporation, hydrogel biophysical properties, and biological outcome. Laminin was efficiently incorporated for all the conditions tested (laminin incorporation > 95%), and the developed hydrogels revealed mechanical properties (average storage modulus (G’) ranging from 187 to 256 Pa) within the values preferred for NSC proliferation and neurite branching and extension. Affinity-bound laminin PEG-4MAL hydrogels better preserve laminin bioactivity, compared to unmodified hydrogels and hydrogels containing physically entrapped laminin, being this effect dependent on NtA concentration. This was evidenced by the 10 M NtA-functionalized PEG-4MAL gels incorporating laminin that support enhanced human NSC proliferation and neurite extension, compared to the latter. Overall, this work highlights the potential of the proposed engineered matrices to be used as defined 3D platforms for the establishment of artificial NSC niches and as extracellular matrix-mimetic microenvironments to support human NSC transplantation. |
id |
RCAP_4aa22e5739730dec6324bab6a68b1320 |
---|---|
oai_identifier_str |
oai:repositorio-aberto.up.pt:10216/122944 |
network_acronym_str |
RCAP |
network_name_str |
Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
repository_id_str |
7160 |
spelling |
Engineering Hydrogels with Affinity-Bound Laminin as 3D Neural Stem Cell Culture SystemsLamininAffinity-bindingHydrogelStem cell nicheNeural stem cellsLaminin incorporation into biological or synthetic hydrogels has been explored to recapitulate the dynamic nature and biological complexity of neural stem cell (NSC) niches. However, the strategies currently explored for laminin immobilization within three-dimensional (3D) matrices do not address a critical aspect influencing cell-matrix interactions, which is the control over laminin conformation and orientation upon immobilization. This is a key feature for the preservation of the protein bioactivity. In this work, we explored an affinity-based approach to mediate the site-selective immobilization of laminin into a degradable synthetic hydrogel. Specifically, a four-arm maleimide terminated poly(ethylene glycol) (PEG-4MAL) macromer was functionalized with a mono-PEGylated recombinant human N-terminal agrin (NtA) domain, to promote high affinity binding of laminin. Different NtA concentrations (10, 50 and 100 M) were used to investigate the impact of NtA density on laminin incorporation, hydrogel biophysical properties, and biological outcome. Laminin was efficiently incorporated for all the conditions tested (laminin incorporation > 95%), and the developed hydrogels revealed mechanical properties (average storage modulus (G’) ranging from 187 to 256 Pa) within the values preferred for NSC proliferation and neurite branching and extension. Affinity-bound laminin PEG-4MAL hydrogels better preserve laminin bioactivity, compared to unmodified hydrogels and hydrogels containing physically entrapped laminin, being this effect dependent on NtA concentration. This was evidenced by the 10 M NtA-functionalized PEG-4MAL gels incorporating laminin that support enhanced human NSC proliferation and neurite extension, compared to the latter. Overall, this work highlights the potential of the proposed engineered matrices to be used as defined 3D platforms for the establishment of artificial NSC niches and as extracellular matrix-mimetic microenvironments to support human NSC transplantation.Royal Society of Chemistry2019-10-122019-10-12T00:00:00Z2020-10-12T00:00:00Zinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleapplication/pdfapplication/pdfhttps://hdl.handle.net/10216/122944eng2047-483010.1039/C9BM00348GBarros, DConde-Sousa, EGonçalves, AMHan, WMGarcía, AJAmaral, IFPêgo, APinfo:eu-repo/semantics/embargoedAccessreponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãoinstacron:RCAAP2023-11-29T14:54:48Zoai:repositorio-aberto.up.pt:10216/122944Portal AgregadorONGhttps://www.rcaap.pt/oai/openaireopendoar:71602024-03-20T00:11:26.143098Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãofalse |
dc.title.none.fl_str_mv |
Engineering Hydrogels with Affinity-Bound Laminin as 3D Neural Stem Cell Culture Systems |
title |
Engineering Hydrogels with Affinity-Bound Laminin as 3D Neural Stem Cell Culture Systems |
spellingShingle |
Engineering Hydrogels with Affinity-Bound Laminin as 3D Neural Stem Cell Culture Systems Barros, D Laminin Affinity-binding Hydrogel Stem cell niche Neural stem cells |
title_short |
Engineering Hydrogels with Affinity-Bound Laminin as 3D Neural Stem Cell Culture Systems |
title_full |
Engineering Hydrogels with Affinity-Bound Laminin as 3D Neural Stem Cell Culture Systems |
title_fullStr |
Engineering Hydrogels with Affinity-Bound Laminin as 3D Neural Stem Cell Culture Systems |
title_full_unstemmed |
Engineering Hydrogels with Affinity-Bound Laminin as 3D Neural Stem Cell Culture Systems |
title_sort |
Engineering Hydrogels with Affinity-Bound Laminin as 3D Neural Stem Cell Culture Systems |
author |
Barros, D |
author_facet |
Barros, D Conde-Sousa, E Gonçalves, AM Han, WM García, AJ Amaral, IF Pêgo, AP |
author_role |
author |
author2 |
Conde-Sousa, E Gonçalves, AM Han, WM García, AJ Amaral, IF Pêgo, AP |
author2_role |
author author author author author author |
dc.contributor.author.fl_str_mv |
Barros, D Conde-Sousa, E Gonçalves, AM Han, WM García, AJ Amaral, IF Pêgo, AP |
dc.subject.por.fl_str_mv |
Laminin Affinity-binding Hydrogel Stem cell niche Neural stem cells |
topic |
Laminin Affinity-binding Hydrogel Stem cell niche Neural stem cells |
description |
Laminin incorporation into biological or synthetic hydrogels has been explored to recapitulate the dynamic nature and biological complexity of neural stem cell (NSC) niches. However, the strategies currently explored for laminin immobilization within three-dimensional (3D) matrices do not address a critical aspect influencing cell-matrix interactions, which is the control over laminin conformation and orientation upon immobilization. This is a key feature for the preservation of the protein bioactivity. In this work, we explored an affinity-based approach to mediate the site-selective immobilization of laminin into a degradable synthetic hydrogel. Specifically, a four-arm maleimide terminated poly(ethylene glycol) (PEG-4MAL) macromer was functionalized with a mono-PEGylated recombinant human N-terminal agrin (NtA) domain, to promote high affinity binding of laminin. Different NtA concentrations (10, 50 and 100 M) were used to investigate the impact of NtA density on laminin incorporation, hydrogel biophysical properties, and biological outcome. Laminin was efficiently incorporated for all the conditions tested (laminin incorporation > 95%), and the developed hydrogels revealed mechanical properties (average storage modulus (G’) ranging from 187 to 256 Pa) within the values preferred for NSC proliferation and neurite branching and extension. Affinity-bound laminin PEG-4MAL hydrogels better preserve laminin bioactivity, compared to unmodified hydrogels and hydrogels containing physically entrapped laminin, being this effect dependent on NtA concentration. This was evidenced by the 10 M NtA-functionalized PEG-4MAL gels incorporating laminin that support enhanced human NSC proliferation and neurite extension, compared to the latter. Overall, this work highlights the potential of the proposed engineered matrices to be used as defined 3D platforms for the establishment of artificial NSC niches and as extracellular matrix-mimetic microenvironments to support human NSC transplantation. |
publishDate |
2019 |
dc.date.none.fl_str_mv |
2019-10-12 2019-10-12T00:00:00Z 2020-10-12T00:00:00Z |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/article |
format |
article |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
https://hdl.handle.net/10216/122944 |
url |
https://hdl.handle.net/10216/122944 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
2047-4830 10.1039/C9BM00348G |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/embargoedAccess |
eu_rights_str_mv |
embargoedAccess |
dc.format.none.fl_str_mv |
application/pdf application/pdf |
dc.publisher.none.fl_str_mv |
Royal Society of Chemistry |
publisher.none.fl_str_mv |
Royal Society of Chemistry |
dc.source.none.fl_str_mv |
reponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação instacron:RCAAP |
instname_str |
Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação |
instacron_str |
RCAAP |
institution |
RCAAP |
reponame_str |
Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
collection |
Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
repository.name.fl_str_mv |
Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação |
repository.mail.fl_str_mv |
|
_version_ |
1799136037220384769 |