{"id":{"repo_id":"toronto-retro","oai_identifier":"oai:utoronto.scholaris.ca:1807/106714"},"canonical_url":"https://search.dev.ndltd.org/etd/toronto-retro/oai:utoronto.scholaris.ca:1807/106714","repository":{"repo_id":"toronto-retro","name":"University of Toronto","base_url":"https://utoronto.scholaris.ca/server/oai/request"},"display":{"title":"Three-dimensional Hyaluronan-based Hydrogels Designed for Breast Cancer Spheroid Formation","abstract":"In vitro culture of cancer cells is traditionally performed with two-dimensional (2D) culture, which fails to model the in vivo tumour cell phenotype and microenvironment. Hyaluronan (HA) is upregulated in breast cancer stroma and thus an excellent starting material in which to culture breast cancer cells; however, HA itself is a viscous liquid. To create a 3D gel for the prolonged culture of breast cancer cells we invented a crosslinked hyaluronan hydrogel comprised of HA-aldehyde and HA-ketone crosslinked with poly(ethylene glycol)-oxyamine. In designing this hydrogel, we considered hydrogel stability, transparency, and mechanical and chemical tunability. These HA-oxime crosslinked gels enabled the long-term culture of breast cancer spheroids (CS). First, hydrogel stiffness was decoupled from the immobilization of cell-adhesive peptides using orthogonal oxime and Diels-Alder click chemistry. Breast CS formation on bifunctional HA hydrogels is dependent on hydrogel stiffness and immobilized peptide. Breast cancer cells treated with the chemotherapeutic doxorubicin were less sensitive compared to cells grown on Matrigel™, a complex protein extract without HA. Breast CS expressed multidrug resistance protein 1, a drug efflux transporter on bifunctional HA hydrogels, and showed decreased doxorubicin drug penetration. Secondly, HA-oxime hydrogels were designed with tunable gelation to encapsulate breast cancer cells. We compared the culture of 5 different cell lines in vitro to in vivo xenografts in mice, thereby benchmarking the gene profile of in vitro cultured cells in our HA-oxime hydrogels to xenografts and relative to more traditional culture strategies of 2D and in Matrigel™. Using these gene expression profiles, we probed drug sensitivity and demonstrated greater predictability in HA-oxime hydrogels than other strategies. Moreover, we demonstrated the capacity to grow patient-derived cells in these HA-oxime hydrogels, paving the way for future opportunities in personalized medicine. Collectively, in these studies we developed and benchmarked in vitro breast cancer models and demonstrate the importance of cell microenvironment when investigating chemotherapeutics.","abstract_html":"In vitro culture of cancer cells is traditionally performed with two-dimensional (2D) culture, which fails to model the in vivo tumour cell phenotype and microenvironment. Hyaluronan (HA) is upregulated in breast cancer stroma and thus an excellent starting material in which to culture breast cancer cells; however, HA itself is a viscous liquid. To create a 3D gel for the prolonged culture of breast cancer cells we invented a crosslinked hyaluronan hydrogel comprised of HA-aldehyde and HA-ketone crosslinked with poly(ethylene glycol)-oxyamine. In designing this hydrogel, we considered hydrogel stability, transparency, and mechanical and chemical tunability. These HA-oxime crosslinked gels enabled the long-term culture of breast cancer spheroids (CS). First, hydrogel stiffness was decoupled from the immobilization of cell-adhesive peptides using orthogonal oxime and Diels-Alder click chemistry. Breast CS formation on bifunctional HA hydrogels is dependent on hydrogel stiffness and immobilized peptide. Breast cancer cells treated with the chemotherapeutic doxorubicin were less sensitive compared to cells grown on Matrigel™, a complex protein extract without HA. Breast CS expressed multidrug resistance protein 1, a drug efflux transporter on bifunctional HA hydrogels, and showed decreased doxorubicin drug penetration. Secondly, HA-oxime hydrogels were designed with tunable gelation to encapsulate breast cancer cells. We compared the culture of 5 different cell lines in vitro to in vivo xenografts in mice, thereby benchmarking the gene profile of in vitro cultured cells in our HA-oxime hydrogels to xenografts and relative to more traditional culture strategies of 2D and in Matrigel™. Using these gene expression profiles, we probed drug sensitivity and demonstrated greater predictability in HA-oxime hydrogels than other strategies. Moreover, we demonstrated the capacity to grow patient-derived cells in these HA-oxime hydrogels, paving the way for future opportunities in personalized medicine. Collectively, in these studies we developed and benchmarked in vitro breast cancer models and demonstrate the importance of cell microenvironment when investigating chemotherapeutics.","abstract_has_math":false,"creators":["Baker, Alexander Edgar Gilbert"],"institution":null,"degree_name":null,"degree_level":null,"degree_discipline":null,"degree_department":"Chemical Engineering Applied Chemistry","school":null,"contributors":[],"advisors":["Shoichet, Molly S"],"committee_chairs":[],"committee_members":[],"year":2019,"date_issued":"2019-06","date_published":"2019-06","updated_at":"2026-07-27T21:28:01Z","subjects":["Biomaterials","Cancer","Drug Screening","Hyaluronan","Hydrogel","Oxime"],"languages":[],"rights":[],"rights_urls":[],"identifier_entries":[]},"links":{"outbound_url":"http://hdl.handle.net/1807/106714","outbound_label":"Handle","outbound_source":"dc:identifier.uri"},"metadata_groups":[{"id":"people","label":"People","entries":[{"key":"dc:contributor.advisor","label":"Advisor","values":["Shoichet, Molly S"]},{"key":"dc:contributor.department","label":"Department","values":["Chemical Engineering Applied Chemistry"]},{"key":"dc:creator","label":"Author","values":["Baker, Alexander Edgar Gilbert"]}]},{"id":"academic_context","label":"Academic Context","entries":[{"key":"dc:date","label":"Dc Date","values":["2019-06"]},{"key":"dc:date.accessioned","label":"Dc Date Accessioned","values":["2021-07-17T04:00:10Z"]},{"key":"dc:date.available","label":"Dc Date Available","values":["2021-07-17T04:00:10Z"]},{"key":"dc:date.issued","label":"Date","values":["2019-06"]},{"key":"dc:type","label":"Dc Type","values":["Thesis"]}]},{"id":"subjects_keywords","label":"Subjects and Keywords","entries":[{"key":"dc:subject","label":"Dc Subject","values":["Biomaterials","Cancer","Drug Screening","Hyaluronan","Hydrogel","Oxime"]}]},{"id":"identifiers","label":"Identifiers","entries":[{"key":"dc:identifier.uri","label":"Identifier URI","values":["http://hdl.handle.net/1807/106714"]}]},{"id":"additional","label":"Additional Metadata","entries":[{"key":"dc:description.abstract","label":"Abstract","values":["In vitro culture of cancer cells is traditionally performed with two-dimensional (2D) culture, which fails to model the in vivo tumour cell phenotype and microenvironment. Hyaluronan (HA) is upregulated in breast cancer stroma and thus an excellent starting material in which to culture breast cancer cells; however, HA itself is a viscous liquid. To create a 3D gel for the prolonged culture of breast cancer cells we invented a crosslinked hyaluronan hydrogel comprised of HA-aldehyde and HA-ketone crosslinked with poly(ethylene glycol)-oxyamine. In designing this hydrogel, we considered hydrogel stability, transparency, and mechanical and chemical tunability. These HA-oxime crosslinked gels enabled the long-term culture of breast cancer spheroids (CS). First, hydrogel stiffness was decoupled from the immobilization of cell-adhesive peptides using orthogonal oxime and Diels-Alder click chemistry. Breast CS formation on bifunctional HA hydrogels is dependent on hydrogel stiffness and immobilized peptide. Breast cancer cells treated with the chemotherapeutic doxorubicin were less sensitive compared to cells grown on Matrigel™, a complex protein extract without HA. Breast CS expressed multidrug resistance protein 1, a drug efflux transporter on bifunctional HA hydrogels, and showed decreased doxorubicin drug penetration. Secondly, HA-oxime hydrogels were designed with tunable gelation to encapsulate breast cancer cells. We compared the culture of 5 different cell lines in vitro to in vivo xenografts in mice, thereby benchmarking the gene profile of in vitro cultured cells in our HA-oxime hydrogels to xenografts and relative to more traditional culture strategies of 2D and in Matrigel™. Using these gene expression profiles, we probed drug sensitivity and demonstrated greater predictability in HA-oxime hydrogels than other strategies. Moreover, we demonstrated the capacity to grow patient-derived cells in these HA-oxime hydrogels, paving the way for future opportunities in personalized medicine. Collectively, in these studies we developed and benchmarked in vitro breast cancer models and demonstrate the importance of cell microenvironment when investigating chemotherapeutics."]},{"key":"dc:description.degree","label":"Dc Description Degree","values":["Ph.D."]},{"key":"dc:title","label":"Title","values":["Three-dimensional Hyaluronan-based Hydrogels Designed for Breast Cancer Spheroid Formation"]}]}],"canonical_facts":{"dc:contributor.advisor":["Shoichet, Molly S"],"dc:contributor.department":["Chemical Engineering Applied Chemistry"],"dc:creator":["Baker, Alexander Edgar Gilbert"],"dc:date":["2019-06"],"dc:date.accessioned":["2021-07-17T04:00:10Z"],"dc:date.available":["2021-07-17T04:00:10Z"],"dc:date.issued":["2019-06"],"dc:description.abstract":["In vitro culture of cancer cells is traditionally performed with two-dimensional (2D) culture, which fails to model the in vivo tumour cell phenotype and microenvironment. Hyaluronan (HA) is upregulated in breast cancer stroma and thus an excellent starting material in which to culture breast cancer cells; however, HA itself is a viscous liquid. To create a 3D gel for the prolonged culture of breast cancer cells we invented a crosslinked hyaluronan hydrogel comprised of HA-aldehyde and HA-ketone crosslinked with poly(ethylene glycol)-oxyamine. In designing this hydrogel, we considered hydrogel stability, transparency, and mechanical and chemical tunability. These HA-oxime crosslinked gels enabled the long-term culture of breast cancer spheroids (CS). First, hydrogel stiffness was decoupled from the immobilization of cell-adhesive peptides using orthogonal oxime and Diels-Alder click chemistry. Breast CS formation on bifunctional HA hydrogels is dependent on hydrogel stiffness and immobilized peptide. Breast cancer cells treated with the chemotherapeutic doxorubicin were less sensitive compared to cells grown on Matrigel™, a complex protein extract without HA. Breast CS expressed multidrug resistance protein 1, a drug efflux transporter on bifunctional HA hydrogels, and showed decreased doxorubicin drug penetration. Secondly, HA-oxime hydrogels were designed with tunable gelation to encapsulate breast cancer cells. We compared the culture of 5 different cell lines in vitro to in vivo xenografts in mice, thereby benchmarking the gene profile of in vitro cultured cells in our HA-oxime hydrogels to xenografts and relative to more traditional culture strategies of 2D and in Matrigel™. Using these gene expression profiles, we probed drug sensitivity and demonstrated greater predictability in HA-oxime hydrogels than other strategies. Moreover, we demonstrated the capacity to grow patient-derived cells in these HA-oxime hydrogels, paving the way for future opportunities in personalized medicine. Collectively, in these studies we developed and benchmarked in vitro breast cancer models and demonstrate the importance of cell microenvironment when investigating chemotherapeutics."],"dc:description.degree":["Ph.D."],"dc:identifier.uri":["http://hdl.handle.net/1807/106714"],"dc:subject":["Biomaterials","Cancer","Drug Screening","Hyaluronan","Hydrogel","Oxime"],"dc:title":["Three-dimensional Hyaluronan-based Hydrogels Designed for Breast Cancer Spheroid Formation"],"dc:type":["Thesis"]},"updated_at":"2026-07-27T21:28:01Z"}