A well plate-based multiplexed platform for incorporation of organoids into an organ-on-a-chip system with a perfusable vasculature.


Journal

Nature protocols
ISSN: 1750-2799
Titre abrégé: Nat Protoc
Pays: England
ID NLM: 101284307

Informations de publication

Date de publication:
04 2021
Historique:
received: 29 09 2020
accepted: 18 12 2020
pubmed: 2 4 2021
medline: 5 5 2021
entrez: 1 4 2021
Statut: ppublish

Résumé

Owing to their high spatiotemporal precision and adaptability to different host cells, organ-on-a-chip systems are showing great promise in drug discovery, developmental biology studies and disease modeling. However, many current micro-engineered biomimetic systems are limited in technological application because of culture media mixing that does not allow direct incorporation of techniques from stem cell biology, such as organoids. Here, we describe a detailed alternative method to cultivate millimeter-scale functional vascularized tissues on a biofabricated platform, termed 'integrated vasculature for assessing dynamic events', that enables facile incorporation of organoid technology. Utilizing the 3D stamping technique with a synthetic polymeric elastomer, a scaffold termed 'AngioTube' is generated with a central microchannel that has the mechanical stability to support a perfusable vascular system and the self-assembly of various parenchymal tissues. We demonstrate an increase in user familiarity and content analysis by situating the scaffold on a footprint of a 96-well plate. Uniquely, the platform can be used for facile connection of two or more tissue compartments in series through a common vasculature. Built-in micropores enable the studies of cell invasion involved in both angiogenesis and metastasis. We describe how this protocol can be applied to create both vascularized cardiac and hepatic tissues, metastatic breast cancer tissue and personalized pancreatic cancer tissue through incorporation of patient-derived organoids. Platform assembly to populating the scaffold with cells of interest into perfusable functional vascularized tissue will require 12-14 d and an additional 4 d if pre-polymer and master molds are needed.

Identifiants

pubmed: 33790475
doi: 10.1038/s41596-020-00490-1
pii: 10.1038/s41596-020-00490-1
doi:

Types de publication

Journal Article Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't

Langues

eng

Sous-ensembles de citation

IM

Pagination

2158-2189

Subventions

Organisme : CIHR
ID : MOP-126027
Pays : Canada
Organisme : CIHR
ID : MOP-137107
Pays : Canada
Organisme : CIHR
ID : MOP-142382
Pays : Canada
Organisme : CIHR
ID : FDN-167274
Pays : Canada
Organisme : NHLBI NIH HHS
ID : R01 HL076485
Pays : United States

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Auteurs

Benjamin Fook Lun Lai (BFL)

Institute of Biomedical Engineering, University of Toronto, Toronto, Ontario, Canada.

Rick Xing Ze Lu (RXZ)

Institute of Biomedical Engineering, University of Toronto, Toronto, Ontario, Canada.

Locke Davenport Huyer (L)

Institute of Biomedical Engineering, University of Toronto, Toronto, Ontario, Canada.
Department of Chemical Engineering and Applied Chemistry, University of Toronto, Toronto, Ontario, Canada.

Sachiro Kakinoki (S)

Faculty of Chemistry, Materials, and Bioengineering, Kansai University, Osaka, Japan.
Organization for Research and Development of Innovative Science and Technology, Kansai University, Osaka, Japan.

Joshua Yazbeck (J)

Department of Chemical Engineering and Applied Chemistry, University of Toronto, Toronto, Ontario, Canada.

Erika Yan Wang (EY)

Institute of Biomedical Engineering, University of Toronto, Toronto, Ontario, Canada.

Qinghua Wu (Q)

Institute of Biomedical Engineering, University of Toronto, Toronto, Ontario, Canada.

Boyang Zhang (B)

Department of Chemical Engineering, McMaster University, Hamilton, Ontario, Canada.

Milica Radisic (M)

Institute of Biomedical Engineering, University of Toronto, Toronto, Ontario, Canada. m.radisic@utoronto.ca.
Department of Chemical Engineering and Applied Chemistry, University of Toronto, Toronto, Ontario, Canada. m.radisic@utoronto.ca.
Toronto General Research Institute, University Health Network, Toronto, Ontario, Canada. m.radisic@utoronto.ca.

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