Abstract
3D organoids using stem cells to study development and disease are now widespread. These models are powerful to mimic in vivo situations but are currently associated with high variability and low throughput. For biomedical research, platforms are thus necessary to increase reproducibility and allow high-throughput screens (HTS). Here, we introduce a microwell platform, integrated in standard culture plates, for functional HTS. Using micro-thermoforming, we form round-bottom microwell arrays from optically clear cyclic olefin polymer films, and assemble them with bottom-less 96-well plates. We show that embryonic stem cells aggregate faster and more reproducibly (centricity, circularity) as compared to a state-of-the-art microwell array. We then run a screen of a chemical library to direct differentiation into primitive endoderm (PrE) and, using on-chip high content imaging (HCI), we identify molecules, including regulators of the cAMP pathway, regulating tissue size, morphology and PrE gene activity. We propose that this platform will benefit to the systematic study of organogenesis in vitro.
References
Oct 1, 1989·Proceedings of the National Academy of Sciences of the United States of America·J F TolesJ E Barker
Apr 12, 2002·Biotechnology and Bioengineering·Stephen M DangPeter W Zandstra
Apr 16, 2005·Blood·Elias T ZambidisCurt I Civin
Feb 16, 2006·IEE Proceedings. Nanobiotechnology·S GiselbrechtK F Weibezahn
May 24, 2006·Biomedical Microdevices·S GiselbrechtA Welle
Feb 3, 2007·BioTechniques·Michael R LamprechtAnne E Carpenter
Jun 1, 2007·Lab on a Chip·Eric GottwaldKarl-Friedrich Weibezahn
May 24, 2008·Nature·Qi-Long YingAustin Smith
Jun 10, 2008·Assay and Drug Development Technologies·Walter D Niles, Peter J Coassin
Aug 30, 2008·Development·Berenika PlusaAnna-Katerina Hadjantonakis
Sep 27, 2008·Lab on a Chip·R TruckenmüllerA Welle
Feb 25, 2010·Development·Marios P StavridisKate G Storey
Mar 9, 2010·Biomaterials·Yoon Young ChoiSang-Hoon Lee
Sep 10, 2010·Development·Jérôme ArtusAnna-Katerina Hadjantonakis
Mar 15, 2011·Advanced Materials·Roman TruckenmüllerClemens van Blitterswijk
Apr 9, 2011·Nature·Mototsugu EirakuYoshiki Sasai
Nov 4, 2011·Biomedical Microdevices·Roman TruckenmüllerDimitrios Stamatialis
Nov 10, 2011·Biomedical Microdevices·Brigitte AltmannAndreas Schober
Jan 19, 2012·British Journal of Pharmacology·Joost C M UitdehaagGuido J R Zaman
Apr 19, 2012·Proceedings of the National Academy of Sciences of the United States of America·Nicolas C RivronClemens A van Blitterswijk
Jan 26, 2013·Science·Martin A Schwartz, Christopher S Chen
May 16, 2013·Nature Communications·K R StevensS N Bhatia
Jun 6, 2013·PloS One·Mijke BuitingaMarcel Karperien
Sep 3, 2013·Nature·Madeline A LancasterJuergen A Knoblich
Sep 27, 2013·Development·Néstor SaizBerenika Plusa
Dec 18, 2013·Nature Cell Biology·M TakasatoM H Little
Apr 3, 2014·PloS One·Adithya SridharSéverine Le Gac
May 8, 2014·Lab on a Chip·K KwapiszewskaZ Brzózka
Sep 23, 2014·Nature Reviews. Molecular Cell Biology·Eliah R Shamir, Andrew J Ewald
Nov 5, 2014·Nature·Kyle W McCrackenJames M Wells
May 11, 2015·Cell·Marc van de WeteringHans Clevers
May 26, 2015·Nature Methods·Anca M PaşcaSergiu P Paşca
Citations
Jun 9, 2016·Expert Opinion on Drug Discovery·Arvind ChandrasekaranChristopher Moraes
Feb 6, 2017·Acta Biomaterialia·Shivem B Shah, Ankur Singh
Apr 8, 2017·Current Opinion in Biotechnology·Tina TronserPavel A Levkin
May 10, 2016·Integrative Biology : Quantitative Biosciences From Nano to Macro·E L Jackson, H Lu
Apr 6, 2018·Lab on a Chip·Jochen KieningerGerald A Urban
Sep 28, 2017·Lab on a Chip·E L Jackson-HolmesH Lu
Sep 9, 2017·Biomedical Microdevices·David BarataSabine van Rijt
Mar 2, 2017·Development·Anna Kicheva, Nicolas C Rivron
Jul 13, 2018·Lab on a Chip·Younggyun LeeNoo Li Jeon
Aug 30, 2018·Electrophoresis·Yue SunXingzhong Zhao
Feb 16, 2019·Advanced Materials·Pinak SamalStefan Giselbrecht
Jul 28, 2019·Current Opinion in Organ Transplantation·Ivan MartinNicolas C Rivron
Nov 7, 2019·World Journal of Stem Cells·Jeong Ah KimWon Jong Rhee
Sep 30, 2017·Scientific Reports·Raja K VadiveluNam-Trung Nguyen
Oct 28, 2017·Frontiers in Pharmacology·Sissi DolciIlaria Decimo
Jul 12, 2016·Molecules : a Journal of Synthetic Chemistry and Natural Product Chemistry·Huei-Wen WuChia-Hsien Hsu
Jul 13, 2016·Cytometry. Part a : the Journal of the International Society for Analytical Cytology·Janos Kriston-Vizi, Horst Flotow
May 4, 2018·Nature·Nicolas C RivronNiels Geijsen
Jan 25, 2021·Acta Biomaterialia·Plansky Hoang, Zhen Ma
Apr 13, 2020·Methods : a Companion to Methods in Enzymology·Steven Vermeulen, Jan de Boer
Nov 6, 2020·Developmental Biology·Nicola GrittiVikas Trivedi
Dec 5, 2017·Materials Science & Engineering. R, Reports : a Review Journal·Jeroen LeijtenAli Khademhosseini
Nov 12, 2017·Trends in Biotechnology·Ross A MarkleinSteven R Bauer
May 26, 2021·Advanced Healthcare Materials·Alexander P M GuttenplanPamela Habibović
May 25, 2021·Biofabrication·Dohui KimDong Sung Kim
May 15, 2020·Chemical Reviews·Carlos MotaLorenzo Moroni
Sep 9, 2021·Development·Nicola GrittiVikas Trivedi