2021
DOI: 10.1126/sciadv.abc1323
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Three-dimensional microscale hanging drop arrays with geometric control for drug screening and live tissue imaging

Abstract: Existing three-dimensional (3D) culture techniques are limited by trade-offs between throughput, capacity for high-resolution imaging in living state, and geometric control. Here, we introduce a modular microscale hanging drop culture where simple design elements allow high replicates for drug screening, direct on-chip real-time or high-resolution confocal microscopy, and geometric control in 3D. Thousands of spheroids can be formed on our microchip in a single step and without any selective pressure from spec… Show more

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Cited by 46 publications
(36 citation statements)
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“…These models are therefore limited to short-term culture and require frequent attention. Improvements to the efficiency and reproducibility of this method include development of hanging drop arrays for use with liquid handling robotics and single cell seeding in nanoliter-sized wells in a microchip format ( Raghavan et al, 2015 ; Ganguli et al, 2021 ). Creation of an open-source, 3D printable multi-purpose hanging drop “dripper” for use with standard tissue culture plates enables metastasis and migration assays as well as co-cultures of cells within the same plate ( Zhao L. et al, 2019 ).…”
Section: 3d In Vitro Ovarian Cancer Modelsmentioning
confidence: 99%
“…These models are therefore limited to short-term culture and require frequent attention. Improvements to the efficiency and reproducibility of this method include development of hanging drop arrays for use with liquid handling robotics and single cell seeding in nanoliter-sized wells in a microchip format ( Raghavan et al, 2015 ; Ganguli et al, 2021 ). Creation of an open-source, 3D printable multi-purpose hanging drop “dripper” for use with standard tissue culture plates enables metastasis and migration assays as well as co-cultures of cells within the same plate ( Zhao L. et al, 2019 ).…”
Section: 3d In Vitro Ovarian Cancer Modelsmentioning
confidence: 99%
“…In conventional hanging drop methods, the small working volume (20–40 μL) on the lid of a plastic plate or in different configuration settings has been a major impediment for culture growth because of the rapid evaporative loss of the small volume of the fluid ( Figure 1E ) ( Tung et al, 2011 ; Klingelhutz et al, 2018 ; Zhao et al, 2019 ; Ganguli et al, 2021 ). In our conceptualized approach, it is possible to use a deep well-plate with up to 1 ml volume in each well to generate pendant drop meniscus, even with the increased capillary length (L c ) of the well depth.…”
Section: Resultsmentioning
confidence: 99%
“…, handing drop insertion ( Tung et al, 2011 ; Zhao et al, 2019 ; Fu et al, 2021 ; Liu et al, 2021 ), including magnetic levitation ( Kim et al, 2013 ; Tseng et al, 2015 ; Ge et al, 2018 ), and nonadhesive surface ( Friedrich et al, 2009 ; Liao et al, 2019 )] were developed to address several applications. Products such as the ultra-low attachment (ULA) plates are available ( Vinci et al, 2012 ; Raghavan et al, 2016 ) along with other techniques for 3D spheroidal culture development, for example, microfabrication ( Lee J. M et al, 2018 ; Park et al, 2018 ; Ganguli et al, 2021 ; Sun et al, 2021 ), soft lithography ( Cha et al, 2017 ; Kuo et al, 2017 ), and floating methods ( Vadivelu et al, 2015 ; Kim et al, 2021 ) among others. Despite the widespread interest, the current culture methods have several limitations, including nonuniformity of spheroid formation, irregular sphericity, short-term cultures, evaporative loss of culture media, and limited volume (size of spheroids) of culture ( Shi et al, 2018 ; De Souza Moraes et al, 2020 ; Han et al, 2021 ).…”
Section: Introductionmentioning
confidence: 99%
“…Three-dimensional tumor spheroids of MCF-7 cells were prepared using hanging drop method, as previous described [ 54 ]. Briefly, 200 μL of agarose gel solution (2%, w/v) were heated at 80 °C and added to a 48-well plate.…”
Section: Methodsmentioning
confidence: 99%