Droplet contact-based spheroid transfer technique as a multi-step assay tool for spheroid arrays

Kim, Hwisoo; Roh, Hyewon; Kim, Haseong; Park, Je‐Kyun

doi:10.1039/d1lc00581b

Cited by 6 publications

(5 citation statements)

References 45 publications

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“…With the help of the gravitational force, the spheroids positioned on top of the pillar structure are transferred into a fresh new medium drop. The underlying working principle of the improved DCST method using the DAC with alignment stopper holes and the PAC with alignment stoppers is similar to that of the previously reported method [ 24 ].…”

Section: Resultsmentioning

confidence: 62%

“…The pillars are also distanced away from each other by 4.5 mm so that it aligns with the DAC. The top of the pillar has a concave plateau structure with air holes for a consistent amount of drop to be transferred onto the pillar and to prevent bubbles from being trapped within the pillar structure [ 24 ]. The dimension of the entire chip is 52 mm × 26 mm in length and width.…”

Section: Resultsmentioning

confidence: 99%

“…Few studies were reported to overcome the labor-intensive and time-consuming problem of handling the spheroids by constructing agarose microwells with slopes of different angles to exchange the medium without damaging the spheroids [ 20 ]. Our group has also previously reported a novel droplet contact-based spheroid transfer (DCST) technique and array chips for simultaneous collection and transplantation of spheroids [ 23 , 24 ]. Despite research, the problem was not resolved as it was still difficult to remove the old cell medium using the slanted microwells while not contacting the spheroids with a pipette tip.…”

Section: Introductionmentioning

confidence: 99%

“…On this basis, this study presents an improved DCST method that simply and uniformly constructs an array of fibroblast-associated glioblastoma multiform spheroid models. Four sets of alignment stoppers have been developed and integrated into the original DCST method previously introduced [ 23 , 24 ] to culture and embed spheroids into hydrogel with perfect alignment. The performance of the improved DCST method with alignment stoppers is compared to that of the original DCST and conventional manual pipetting methods in the areas of spheroid shape, retention rate, and uniformity of the spheroid models.…”

Section: Introductionmentioning

confidence: 99%

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Construction of a Fibroblast-Associated Tumor Spheroid Model Based on a Collagen Drop Array Chip

2021

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Spheroid, a 3D aggregate of tumor cells in a spherical shape, has overcome the limitations of conventional 3D cell models to accurately mimic the in-vivo environment of a human body. The spheroids are cultured with other primary cells and embedded in collagen drops using hang drop plates and low-attachment well plates to construct a spheroid–hydrogel model that better mimics the cell–cell and cell–extracellular matrix (ECM) interactions. However, the conventional methods of culturing and embedding spheroids into ECM have several shortcomings. The procedure of transferring a single spheroid at a time by manual pipetting results in well-to-well variation and even loss or damage of the spheroid. Based on the previously introduced droplet contact-based spheroid transfer technique, we present a poly(dimethylsiloxane) and resin-based drop array chip and a pillar array chip with alignment stoppers, which enhances the alignment between the chips for uniform placement of spheroids. This method allows the facile and stable transfer of the spheroid array and even eliminates the need for a stereomicroscope while handling the cell models. The novel platform demonstrates a homogeneous and time-efficient construction and diverse analysis of an array of fibroblast-associated glioblastoma multiforme spheroids that are embedded in collagen.

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Section: Resultsmentioning

confidence: 62%

Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Construction of a Fibroblast-Associated Tumor Spheroid Model Based on a Collagen Drop Array Chip

2021

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“…2C). 60 This technique allows the controlled change of reagents of individual spheroids, and therefore gives more experimental control for investigating the effects of a particular drug. 60 The manipulation is, however, limited spatially as spheroid-containing microdroplets can only be transferred between pillars and wells.…”

Section: Hydrodynamic Techniquesmentioning

confidence: 99%

Tools for manipulation and positioning of microtissues

et al. 2022

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Reconfigurable Hanging Drop Microarray Platform for On‐Demand Preparation and Analysis of Spheroid Array

Kim,

Kim

et al. 2024

Adv Healthcare Materials

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In response to the increasing demand for spheroid‐based cancer research, the importance of developing integrated platforms that can simultaneously facilitate high‐throughput spheroid production and multiplexed analysis has been emphasized. In addition, the understanding of how the size and cellular composition of tumors directly influence their internal structures and functionalities underlines the critical need to produce spheroids of diverse sizes and compositions on a large scale. To address this rising demand, we present a configurable and linkable in vitro 3D cell culture kit for spheroids, termed CLiCK‐Spheroid. This platform consists of three primary components: a hanging drop microarray (HDMA), concave pillar microarray (CPMA), and gradient blocks. The HDMA alone produces a homogeneous spheroid array, while its combination with the gradient block enables one‐step generation of a size‐gradient spheroid array. Using the size‐gradient spheroid arrays, the occurrence of necrotic cores based on spheroid size is demonstrated. Additionally, spheroids in a single batch can be conveniently compartmentalized and regrouped using CPMA, enhancing the versatility of spheroid arrays and enabling multiplexed drug treatments. By combining the different assembly methods, we have achieved a high‐throughput production of cell composition‐gradient spheroid arrays, with noticeable variations in morphology and vascularization based on cell compositions.This article is protected by copyright. All rights reserved

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Droplet contact-based spheroid transfer technique as a multi-step assay tool for spheroid arrays

Abstract: Hanging drop plates and low-attachment well plates are suitable for a high throughput screening model of a spheroid, because each drop (or well) contains a single spheroid and the spheroid...

Cited by 6 publications

References 45 publications

Construction of a Fibroblast-Associated Tumor Spheroid Model Based on a Collagen Drop Array Chip

Construction of a Fibroblast-Associated Tumor Spheroid Model Based on a Collagen Drop Array Chip

Tools for manipulation and positioning of microtissues

Reconfigurable Hanging Drop Microarray Platform for On‐Demand Preparation and Analysis of Spheroid Array

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