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Conference Paper: Time-course of endothelial differentiation by self-assembly of dental stem cells

TitleTime-course of endothelial differentiation by self-assembly of dental stem cells
Authors
Issue Date2017
PublisherInternational Association for Dental Research.
Citation
The 31st Annual Scientific Meeting of the International Association for Dental Research Southeast Asian Division (IADR-SEA), 28th Annual Scientific Meeting of South East Asia Association for Dental Education (SEAADE) & 40th Chinese Taipei Association for Dental Sciences, Taipei, Taiwan, 10-13 August 2017, Final Presentation ID: 0145 How to Cite?
AbstractObjectives: The current study aimed to investigate the time-course effects on endothelial differentiation of dental stem cells, when they are allowed to self-assemble into microtissues. Methods: Micro-molds purchased from Microtissues, Inc. were used to fabricate agarose three-dimensional (3D) petri-dishes, which contain circular recesses. Dental pulp stem cells (DPSCs) and stem cells from human exfoliated deciduous teeth (SHED) suspensions were poured into these petri-dishes and allowed to self-assemble into 3D microtissue spheroids over 24 hours. After 24 hours, medium was changed to fully supplemented endothelial growth medium -2 (Lonza Biologics Inc.) and subsequently changed in every three days. After 3-, 6- and 9- days, microtissue spheroids were transferred onto the two-dimensional culture dishes and allowed to attach and dissociate into single cells. At confluence, cells were trypsinized and examined for expression of endothelial markers – vWF, CD31, eNOS, VE-cadherin, VEGFR-1 and 2 via qPCR, western blotting and immunofluorescence. The capacity of these cells to form in-vitro capillary-like tubes on basement membrane matrix – Matrigel was assessed. Results: qPCR and immunofluorescence results showed a significantly higher levels of expression of endothelial markers in 3-days induced, microtissue-derived DPSCs and SHED compared to that of 6- and 9- days induced cells. Accordingly, 3-days induced, microtissue-derived SHED were able to form extensive capillary-like tubes on Matrigel, while cells derived from microtissues induced for 6- and 9- days failed to form an extensive tube-network. In contrast, microtissue-derived DPSCs failed to form capillary-like tubes regardless of the duration of induction. Both DPSCs and SHED, when cultured in traditional two-dimensional cultures did not show a significant increase in endothelial marker expression or capillary-like tube formation on Matrigel regardless of the duration of induction. Conclusions: SHED hold a higher potential for endothelial differentiation, when they were self-assembled into 3D spheroids and induced for 3 days compared to that of DPSCs.
DescriptionOral Session 10 Mineralized Tissue / Pulp and Stem Cell Biology - Final Presentation ID: 0145
Persistent Identifierhttp://hdl.handle.net/10722/308261

 

DC FieldValueLanguage
dc.contributor.authorDissanayaka, WL-
dc.contributor.authorZhang, C-
dc.date.accessioned2021-11-12T13:44:45Z-
dc.date.available2021-11-12T13:44:45Z-
dc.date.issued2017-
dc.identifier.citationThe 31st Annual Scientific Meeting of the International Association for Dental Research Southeast Asian Division (IADR-SEA), 28th Annual Scientific Meeting of South East Asia Association for Dental Education (SEAADE) & 40th Chinese Taipei Association for Dental Sciences, Taipei, Taiwan, 10-13 August 2017, Final Presentation ID: 0145-
dc.identifier.urihttp://hdl.handle.net/10722/308261-
dc.descriptionOral Session 10 Mineralized Tissue / Pulp and Stem Cell Biology - Final Presentation ID: 0145-
dc.description.abstractObjectives: The current study aimed to investigate the time-course effects on endothelial differentiation of dental stem cells, when they are allowed to self-assemble into microtissues. Methods: Micro-molds purchased from Microtissues, Inc. were used to fabricate agarose three-dimensional (3D) petri-dishes, which contain circular recesses. Dental pulp stem cells (DPSCs) and stem cells from human exfoliated deciduous teeth (SHED) suspensions were poured into these petri-dishes and allowed to self-assemble into 3D microtissue spheroids over 24 hours. After 24 hours, medium was changed to fully supplemented endothelial growth medium -2 (Lonza Biologics Inc.) and subsequently changed in every three days. After 3-, 6- and 9- days, microtissue spheroids were transferred onto the two-dimensional culture dishes and allowed to attach and dissociate into single cells. At confluence, cells were trypsinized and examined for expression of endothelial markers – vWF, CD31, eNOS, VE-cadherin, VEGFR-1 and 2 via qPCR, western blotting and immunofluorescence. The capacity of these cells to form in-vitro capillary-like tubes on basement membrane matrix – Matrigel was assessed. Results: qPCR and immunofluorescence results showed a significantly higher levels of expression of endothelial markers in 3-days induced, microtissue-derived DPSCs and SHED compared to that of 6- and 9- days induced cells. Accordingly, 3-days induced, microtissue-derived SHED were able to form extensive capillary-like tubes on Matrigel, while cells derived from microtissues induced for 6- and 9- days failed to form an extensive tube-network. In contrast, microtissue-derived DPSCs failed to form capillary-like tubes regardless of the duration of induction. Both DPSCs and SHED, when cultured in traditional two-dimensional cultures did not show a significant increase in endothelial marker expression or capillary-like tube formation on Matrigel regardless of the duration of induction. Conclusions: SHED hold a higher potential for endothelial differentiation, when they were self-assembled into 3D spheroids and induced for 3 days compared to that of DPSCs.-
dc.languageeng-
dc.publisherInternational Association for Dental Research.-
dc.relation.ispartofIADR-SEA & SEAADE (International Association for Dental Research South East Asian Division Meeting), 2017-
dc.titleTime-course of endothelial differentiation by self-assembly of dental stem cells-
dc.typeConference_Paper-
dc.identifier.emailDissanayaka, WL: warunad@hku.hk-
dc.identifier.emailZhang, C: zhangcf@hku.hk-
dc.identifier.authorityDissanayaka, WL=rp02216-
dc.identifier.authorityZhang, C=rp01408-
dc.identifier.hkuros329489-
dc.identifier.spageFinal Presentation ID: 0145-
dc.identifier.epageFinal Presentation ID: 0145-

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