Enzyme-mediated hyaluronic acid–tyramine hydrogels for the propagation of human embryonic stem cells in 3D
[Display omitted] The propagation of human embryonic stem cells (hESCs) in three-dimensional (3D) scaffolds facilitates the cell expansion process and supplies pluripotent cells of high quality for broad-spectrum applications in regenerative medicine. Herein, we report an enzyme-mediated hyaluronic...
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| Veröffentlicht in: | Acta biomaterialia 2015-09, Vol.24, p.159-171 |
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| creator | Xu, Keming Narayanan, Karthikeyan Lee, Fan Bae, Ki Hyun Gao, Shujun Kurisawa, Motoichi |
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The propagation of human embryonic stem cells (hESCs) in three-dimensional (3D) scaffolds facilitates the cell expansion process and supplies pluripotent cells of high quality for broad-spectrum applications in regenerative medicine. Herein, we report an enzyme-mediated hyaluronic acid–tyramine (HA–Tyr) hydrogel that encapsulated and propagated hESCs in 3D. HA–Tyr hydrogels were formed by crosslinking the tyramine moieties with horseradish peroxidase (HRP) and hydrogen peroxide (H2O2). By changing the HRP and H2O2 concentration, we prepared HA–Tyr hydrogels of different mechanical strength and studied the self-renewal properties of hESCs in these scaffolds. We observed that both the chemical composition and mechanical strength of substrates were important factors affecting cell proliferation and pluripotency. The HA–Tyr hydrogel with a compressive modulus of ∼350Pa supported the proliferation of hESCs at the pluripotent state in both mTeSR1 medium and mouse embryonic fibroblast (MEF)-conditioned medium. Immunohistochemical analyses revealed that hESCs proliferated well and formed spheroid structures in 3D, without undergoing apoptosis. The hESCs cultured in HA–Tyr hydrogels showed high expression of CD44 and pluripotency markers. These cells exhibited the capability to form cell derivatives of all three embryonic germ layers in vitro and in vivo. In addition, the genetic integrity of the hESCs was unaffected in the 3D cultivation system.
The scope of this study is to provide a stable 3D cultivation system for the expansion of human embryonic stem cells (hESCs) towards clinical applications. We report an enzyme mediated hyaluronic acid–tyramine (HA–Tyr) hydrogel that encapsulated and propagated hESCs in 3D. Unlike other HA-based photo-crosslinked hydrogel systems reported, we investigated the effects of mechanical strength of hydrogels on the self-renewal properties of hESCs in 3D. Then, we characterized hESCs cultured in hydrogels with lower mechanical strength that best supported the self-renewal of hESCs. Hence, we demonstrated a reliable approach for the controlled propagation of hESCs in 3D. We believe that such an approach would facilitate the development of stem cell-based therapy towards clinical applications. |
| doi_str_mv | 10.1016/j.actbio.2015.06.026 |
| format | Article |
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The propagation of human embryonic stem cells (hESCs) in three-dimensional (3D) scaffolds facilitates the cell expansion process and supplies pluripotent cells of high quality for broad-spectrum applications in regenerative medicine. Herein, we report an enzyme-mediated hyaluronic acid–tyramine (HA–Tyr) hydrogel that encapsulated and propagated hESCs in 3D. HA–Tyr hydrogels were formed by crosslinking the tyramine moieties with horseradish peroxidase (HRP) and hydrogen peroxide (H2O2). By changing the HRP and H2O2 concentration, we prepared HA–Tyr hydrogels of different mechanical strength and studied the self-renewal properties of hESCs in these scaffolds. We observed that both the chemical composition and mechanical strength of substrates were important factors affecting cell proliferation and pluripotency. The HA–Tyr hydrogel with a compressive modulus of ∼350Pa supported the proliferation of hESCs at the pluripotent state in both mTeSR1 medium and mouse embryonic fibroblast (MEF)-conditioned medium. Immunohistochemical analyses revealed that hESCs proliferated well and formed spheroid structures in 3D, without undergoing apoptosis. The hESCs cultured in HA–Tyr hydrogels showed high expression of CD44 and pluripotency markers. These cells exhibited the capability to form cell derivatives of all three embryonic germ layers in vitro and in vivo. In addition, the genetic integrity of the hESCs was unaffected in the 3D cultivation system.
The scope of this study is to provide a stable 3D cultivation system for the expansion of human embryonic stem cells (hESCs) towards clinical applications. We report an enzyme mediated hyaluronic acid–tyramine (HA–Tyr) hydrogel that encapsulated and propagated hESCs in 3D. Unlike other HA-based photo-crosslinked hydrogel systems reported, we investigated the effects of mechanical strength of hydrogels on the self-renewal properties of hESCs in 3D. Then, we characterized hESCs cultured in hydrogels with lower mechanical strength that best supported the self-renewal of hESCs. Hence, we demonstrated a reliable approach for the controlled propagation of hESCs in 3D. We believe that such an approach would facilitate the development of stem cell-based therapy towards clinical applications.</description><identifier>ISSN: 1742-7061</identifier><identifier>EISSN: 1878-7568</identifier><identifier>DOI: 10.1016/j.actbio.2015.06.026</identifier><identifier>PMID: 26112373</identifier><language>eng</language><publisher>England: Elsevier Ltd</publisher><subject>3D culture ; Animals ; Cultivation ; Embryonic stem cells ; Encapsulation ; Horseradish Peroxidase - chemistry ; Human ; Human Embryonic Stem Cells - cytology ; Human Embryonic Stem Cells - metabolism ; Humans ; Hyaluronic acid ; Hyaluronic Acid - chemistry ; Hydrogel ; Hydrogels ; Hydrogels - chemistry ; Hydrogen Peroxide - chemistry ; Materials Testing ; Mice ; Scaffold ; Scaffolds ; Stem cells ; Strength ; Three dimensional ; Tissue Scaffolds - chemistry ; Tyramine - chemistry</subject><ispartof>Acta biomaterialia, 2015-09, Vol.24, p.159-171</ispartof><rights>2015 Acta Materialia Inc.</rights><rights>Copyright © 2015 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c531t-44c5689ac079586edc6817656bbe9d75ff26994e0c4b0411c8feade4d099efb3</citedby><cites>FETCH-LOGICAL-c531t-44c5689ac079586edc6817656bbe9d75ff26994e0c4b0411c8feade4d099efb3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://www.sciencedirect.com/science/article/pii/S1742706115002950$$EHTML$$P50$$Gelsevier$$H</linktohtml><link.rule.ids>314,776,780,3537,27901,27902,65306</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/26112373$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Xu, Keming</creatorcontrib><creatorcontrib>Narayanan, Karthikeyan</creatorcontrib><creatorcontrib>Lee, Fan</creatorcontrib><creatorcontrib>Bae, Ki Hyun</creatorcontrib><creatorcontrib>Gao, Shujun</creatorcontrib><creatorcontrib>Kurisawa, Motoichi</creatorcontrib><title>Enzyme-mediated hyaluronic acid–tyramine hydrogels for the propagation of human embryonic stem cells in 3D</title><title>Acta biomaterialia</title><addtitle>Acta Biomater</addtitle><description>[Display omitted]
The propagation of human embryonic stem cells (hESCs) in three-dimensional (3D) scaffolds facilitates the cell expansion process and supplies pluripotent cells of high quality for broad-spectrum applications in regenerative medicine. Herein, we report an enzyme-mediated hyaluronic acid–tyramine (HA–Tyr) hydrogel that encapsulated and propagated hESCs in 3D. HA–Tyr hydrogels were formed by crosslinking the tyramine moieties with horseradish peroxidase (HRP) and hydrogen peroxide (H2O2). By changing the HRP and H2O2 concentration, we prepared HA–Tyr hydrogels of different mechanical strength and studied the self-renewal properties of hESCs in these scaffolds. We observed that both the chemical composition and mechanical strength of substrates were important factors affecting cell proliferation and pluripotency. The HA–Tyr hydrogel with a compressive modulus of ∼350Pa supported the proliferation of hESCs at the pluripotent state in both mTeSR1 medium and mouse embryonic fibroblast (MEF)-conditioned medium. Immunohistochemical analyses revealed that hESCs proliferated well and formed spheroid structures in 3D, without undergoing apoptosis. The hESCs cultured in HA–Tyr hydrogels showed high expression of CD44 and pluripotency markers. These cells exhibited the capability to form cell derivatives of all three embryonic germ layers in vitro and in vivo. In addition, the genetic integrity of the hESCs was unaffected in the 3D cultivation system.
The scope of this study is to provide a stable 3D cultivation system for the expansion of human embryonic stem cells (hESCs) towards clinical applications. We report an enzyme mediated hyaluronic acid–tyramine (HA–Tyr) hydrogel that encapsulated and propagated hESCs in 3D. Unlike other HA-based photo-crosslinked hydrogel systems reported, we investigated the effects of mechanical strength of hydrogels on the self-renewal properties of hESCs in 3D. Then, we characterized hESCs cultured in hydrogels with lower mechanical strength that best supported the self-renewal of hESCs. Hence, we demonstrated a reliable approach for the controlled propagation of hESCs in 3D. We believe that such an approach would facilitate the development of stem cell-based therapy towards clinical applications.</description><subject>3D culture</subject><subject>Animals</subject><subject>Cultivation</subject><subject>Embryonic stem cells</subject><subject>Encapsulation</subject><subject>Horseradish Peroxidase - chemistry</subject><subject>Human</subject><subject>Human Embryonic Stem Cells - cytology</subject><subject>Human Embryonic Stem Cells - metabolism</subject><subject>Humans</subject><subject>Hyaluronic acid</subject><subject>Hyaluronic Acid - chemistry</subject><subject>Hydrogel</subject><subject>Hydrogels</subject><subject>Hydrogels - chemistry</subject><subject>Hydrogen Peroxide - chemistry</subject><subject>Materials Testing</subject><subject>Mice</subject><subject>Scaffold</subject><subject>Scaffolds</subject><subject>Stem cells</subject><subject>Strength</subject><subject>Three dimensional</subject><subject>Tissue Scaffolds - chemistry</subject><subject>Tyramine - chemistry</subject><issn>1742-7061</issn><issn>1878-7568</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2015</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqNUU1r3DAUFCEh3_-gFB1zsSvJtiRfCiVN00Igl70LWXre1WJbG0kuuKf-h_7D_pJqs0mPIaf34M28GWYQ-kBJSQnln7alNqlzvmSENiXhJWH8CJ1TKWQhGi6P8y5qVgjC6Rm6iHFLSCUpk6fojHFKWSWqczTcTb-WEYoRrNMJLN4sepiDn5zB2jj79_eftAQ9ugnyyQa_hiHi3gecNoB3we_0WifnJ-x7vJlHPWEYu7A8P4gJRmxgyAw34errFTrp9RDh-mVeotW3u9Xt9-Lh8f7H7ZeHwjQVTUVdm-y_1YaItpEcrOGSCt7wroPWiqbvGW_bGoipO1JTamQP2kJtSdtC31WX6ObwNtt7miEmNbq4t6En8HNUVFQkh5K13gGlTHApGM_Q-gA1wccYoFe74EYdFkWJ2jeiturQiNo3oghX5Jn28UVh7nLI_0mvFWTA5wMgBws_HQQVjYPJ5EICmKSsd28r_AOR0qCC</recordid><startdate>201509</startdate><enddate>201509</enddate><creator>Xu, Keming</creator><creator>Narayanan, Karthikeyan</creator><creator>Lee, Fan</creator><creator>Bae, Ki Hyun</creator><creator>Gao, Shujun</creator><creator>Kurisawa, Motoichi</creator><general>Elsevier Ltd</general><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7QO</scope><scope>8FD</scope><scope>FR3</scope><scope>P64</scope><scope>7SR</scope><scope>7TB</scope><scope>7U5</scope><scope>8BQ</scope><scope>F28</scope><scope>JG9</scope><scope>L7M</scope></search><sort><creationdate>201509</creationdate><title>Enzyme-mediated hyaluronic acid–tyramine hydrogels for the propagation of human embryonic stem cells in 3D</title><author>Xu, Keming ; Narayanan, Karthikeyan ; Lee, Fan ; Bae, Ki Hyun ; Gao, Shujun ; Kurisawa, Motoichi</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c531t-44c5689ac079586edc6817656bbe9d75ff26994e0c4b0411c8feade4d099efb3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2015</creationdate><topic>3D culture</topic><topic>Animals</topic><topic>Cultivation</topic><topic>Embryonic stem cells</topic><topic>Encapsulation</topic><topic>Horseradish Peroxidase - chemistry</topic><topic>Human</topic><topic>Human Embryonic Stem Cells - cytology</topic><topic>Human Embryonic Stem Cells - metabolism</topic><topic>Humans</topic><topic>Hyaluronic acid</topic><topic>Hyaluronic Acid - chemistry</topic><topic>Hydrogel</topic><topic>Hydrogels</topic><topic>Hydrogels - chemistry</topic><topic>Hydrogen Peroxide - chemistry</topic><topic>Materials Testing</topic><topic>Mice</topic><topic>Scaffold</topic><topic>Scaffolds</topic><topic>Stem cells</topic><topic>Strength</topic><topic>Three dimensional</topic><topic>Tissue Scaffolds - chemistry</topic><topic>Tyramine - chemistry</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Xu, Keming</creatorcontrib><creatorcontrib>Narayanan, Karthikeyan</creatorcontrib><creatorcontrib>Lee, Fan</creatorcontrib><creatorcontrib>Bae, Ki Hyun</creatorcontrib><creatorcontrib>Gao, Shujun</creatorcontrib><creatorcontrib>Kurisawa, Motoichi</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Biotechnology Research Abstracts</collection><collection>Technology Research Database</collection><collection>Engineering Research Database</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>Engineered Materials Abstracts</collection><collection>Mechanical & Transportation Engineering Abstracts</collection><collection>Solid State and Superconductivity Abstracts</collection><collection>METADEX</collection><collection>ANTE: Abstracts in New Technology & Engineering</collection><collection>Materials Research Database</collection><collection>Advanced Technologies Database with Aerospace</collection><jtitle>Acta biomaterialia</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Xu, Keming</au><au>Narayanan, Karthikeyan</au><au>Lee, Fan</au><au>Bae, Ki Hyun</au><au>Gao, Shujun</au><au>Kurisawa, Motoichi</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Enzyme-mediated hyaluronic acid–tyramine hydrogels for the propagation of human embryonic stem cells in 3D</atitle><jtitle>Acta biomaterialia</jtitle><addtitle>Acta Biomater</addtitle><date>2015-09</date><risdate>2015</risdate><volume>24</volume><spage>159</spage><epage>171</epage><pages>159-171</pages><issn>1742-7061</issn><eissn>1878-7568</eissn><abstract>[Display omitted]
The propagation of human embryonic stem cells (hESCs) in three-dimensional (3D) scaffolds facilitates the cell expansion process and supplies pluripotent cells of high quality for broad-spectrum applications in regenerative medicine. Herein, we report an enzyme-mediated hyaluronic acid–tyramine (HA–Tyr) hydrogel that encapsulated and propagated hESCs in 3D. HA–Tyr hydrogels were formed by crosslinking the tyramine moieties with horseradish peroxidase (HRP) and hydrogen peroxide (H2O2). By changing the HRP and H2O2 concentration, we prepared HA–Tyr hydrogels of different mechanical strength and studied the self-renewal properties of hESCs in these scaffolds. We observed that both the chemical composition and mechanical strength of substrates were important factors affecting cell proliferation and pluripotency. The HA–Tyr hydrogel with a compressive modulus of ∼350Pa supported the proliferation of hESCs at the pluripotent state in both mTeSR1 medium and mouse embryonic fibroblast (MEF)-conditioned medium. Immunohistochemical analyses revealed that hESCs proliferated well and formed spheroid structures in 3D, without undergoing apoptosis. The hESCs cultured in HA–Tyr hydrogels showed high expression of CD44 and pluripotency markers. These cells exhibited the capability to form cell derivatives of all three embryonic germ layers in vitro and in vivo. In addition, the genetic integrity of the hESCs was unaffected in the 3D cultivation system.
The scope of this study is to provide a stable 3D cultivation system for the expansion of human embryonic stem cells (hESCs) towards clinical applications. We report an enzyme mediated hyaluronic acid–tyramine (HA–Tyr) hydrogel that encapsulated and propagated hESCs in 3D. Unlike other HA-based photo-crosslinked hydrogel systems reported, we investigated the effects of mechanical strength of hydrogels on the self-renewal properties of hESCs in 3D. Then, we characterized hESCs cultured in hydrogels with lower mechanical strength that best supported the self-renewal of hESCs. Hence, we demonstrated a reliable approach for the controlled propagation of hESCs in 3D. We believe that such an approach would facilitate the development of stem cell-based therapy towards clinical applications.</abstract><cop>England</cop><pub>Elsevier Ltd</pub><pmid>26112373</pmid><doi>10.1016/j.actbio.2015.06.026</doi><tpages>13</tpages></addata></record> |
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| ispartof | Acta biomaterialia, 2015-09, Vol.24, p.159-171 |
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| subjects | 3D culture Animals Cultivation Embryonic stem cells Encapsulation Horseradish Peroxidase - chemistry Human Human Embryonic Stem Cells - cytology Human Embryonic Stem Cells - metabolism Humans Hyaluronic acid Hyaluronic Acid - chemistry Hydrogel Hydrogels Hydrogels - chemistry Hydrogen Peroxide - chemistry Materials Testing Mice Scaffold Scaffolds Stem cells Strength Three dimensional Tissue Scaffolds - chemistry Tyramine - chemistry |
| title | Enzyme-mediated hyaluronic acid–tyramine hydrogels for the propagation of human embryonic stem cells in 3D |
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