Multiwell plate integrated with transparent liquid marbles for 3D cell culturing

Xiao Lin , Mei Duan , Hui Zhang , Haohao Jiang , Heng Liu , Xianglong Pang , Wenjun Tian , Chenxi Yun , Xiaoguang Li

Droplet ›› 2025, Vol. 4 ›› Issue (2) : e164

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Droplet ›› 2025, Vol. 4 ›› Issue (2) :e164 DOI: 10.1002/dro2.164
RESEARCH ARTICLE

Multiwell plate integrated with transparent liquid marbles for 3D cell culturing

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Abstract

The development of 3D cell culturing toward labor saving and versatility is highly desired. Here, we propose a platform consisting of a multiwell plate and liquid marbles (LMs). The inner walls of the plate are covered with particle-detachable superhydrophobic coatings that serve as both the substrates and particle sources for LM production. A produced LM, which serves as a minireactor for the 3D culture, features a monolayer nanoparticulate shell and naked-droplet-like transparency. The LM-in-plate platform provides a double-superhydrophobic environment that increases the stability of the 3D culture and reduces the necessary operational cautions. In addition, both cell observation and high-throughput applications can be conducted in situ, owing to the high LM transparency and the multiwell structure, respectively. This platform integrates the advantages of naked droplets (transparent and clean), LMs (stable non-wetting), and multiwell plates (high-throughput capability) and thus is promising for labor-saving and versatile 3D culturing.

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Xiao Lin, Mei Duan, Hui Zhang, Haohao Jiang, Heng Liu, Xianglong Pang, Wenjun Tian, Chenxi Yun, Xiaoguang Li. Multiwell plate integrated with transparent liquid marbles for 3D cell culturing. Droplet, 2025, 4(2): e164 DOI:10.1002/dro2.164

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References

[1]

Kim J, Koo BK, Knoblich JA. Human organoids: model systems for human biology and medicine. Nat Rev Mol Cell Biol. 2020; 21: 571-584.

[2]

Park J, Lee GH, Yull Park J, Lee JC, Kim HC. Hypergravity-induced multicellular spheroid generation with different morphological patterns precisely controlled on a centrifugal microfluidic platform. Biofabrication. 2017; 9:045006.

[3]

Gallegos-Martinez S, Lara-Mayorga IM, Samandari M, et al. Culture of cancer spheroids and evaluation of anti-cancer drugs in 3D-printed miniaturized continuous stirred tank reactors (mCSTRs). Biofabrication. 2022; 14:035007.

[4]

Rodoplu D, Matahum JS, Hsu CH. A microfluidic hanging drop-based spheroid co-culture platform for probing tumor angiogenesis. Lab Chip. 2022; 22: 1275-1285.

[5]

Oliveira MB, Neto AI, Correia CR, Rial-Hermida MI, Alvarez-Lorenzo C, Mano JF. Superhydrophobic chips for cell spheroids high-throughput generation and drug screening. ACS Appl Mater Interfaces. 2014; 6: 9488-9495.

[6]

Cui H, Tronser T, Wang X, et al. High-throughput formation of miniaturized cocultures of 2D cell monolayers and 3D cell spheroids using droplet microarray. Droplet. 2023; 2:e39.

[7]

Chen M, Shah MP, Shelper TB, et al. Naked liquid marbles: a robust three-dimensional low-volume cell-culturing system. ACS Appl Mater Interfaces. 2019; 11: 9814-9823.

[8]

Xu L, Chen S, Lu X, Lu Q. Durable superamphiphobic silica aerogel surfaces for the culture of 3D cellular spheroids. Natl Sci Rev. 2019; 6: 1255-1265.

[9]

Aussillous P, Quere D. Liquid marbles. Nature. 2001; 411: 924-927.

[10]

Arbatan T, Al-Abboodi A, Sarvi F, Chan PP, Shen W. Tumor inside a pearl drop. Adv Healthc Mater. 2012; 1: 467-469.

[11]

Sarvi F, Arbatan T, Chan PPY, Shen W. A novel technique for the formation of embryoid bodies inside liquid marbles. RSC Adv. 2013; 3: 14501-14508.

[12]

Sarvi F, Jain K, Arbatan T, et al. Cardiogenesis of embryonic stem cells with liquid marble micro-bioreactor. Adv Healthc Mater. 2015; 4: 77-86.

[13]

Lin K, Chen R, Zhang L, Zang D, Geng X, Shen W. Transparent bioreactors based on nanoparticle-coated liquid marbles for in situ observation of suspending embryonic body formation and differentiation. ACS Appl Mater Interfaces. 2019; 11: 8789-8796.

[14]

Vadivelu RK, Ooi CH, Yao R-Q, et al. Generation of three-dimensional multiple spheroid model of olfactory ensheathing cells using floating liquid marbles. Sci Rep. 2015; 5:15083.

[15]

Li H, Liu P, Kaur G, Yao X, Yang M. Transparent and gas-permeable liquid marbles for culturing and drug sensitivity test of tumor spheroids. Adv Healthc Mater. 2017; 6:15215.

[16]

Zhang J, Chintalaramulu N, Vadivelu R, et al. Inertial microfluidic purification of floating cancer cells for drug screening and three-dimensional tumor models. Anal Chem. 2020; 92: 11558-11564.

[17]

Vadivelu R, Kashaninejad N, Nikmaneshi MR, et al. Sessile liquid marbles with embedded hydrogels as bioreactors for three-dimensional cell culture. Adv Biol. 2021; 5:2170022.

[18]

Kroupova J, Hanus J, Stepanek F. Surprising efficacy twist of two established cytostatics revealed by a-la-carte 3D cell spheroid preparation protocol. Eur J Pharm Biopharm. 2022; 180: 224-237.

[19]

Langella A, Gadau SD, Serra E, Bebbere D, Ledda S. Microtubular assessment of C6 rat glioma cell spheroids developed in transparent liquid marbles or hanging drops. Biology (Basel). 2022; 11: 492.

[20]

Fernandez-Montoro A, Angel-Velez D, Benedetti C, et al. Alternative culture systems for bovine oocyte in vitro maturation: liquid marbles and differentially shaped 96-well plates. Animals (Basel). 2023; 13: 1635.

[21]

Aalders J, Leger L, Tuerlings T, Ledda S, van Hengel J. Liquid marble technology to create cost-effective 3D cardiospheres as a platform for in vitro drug testing and disease modelling. Methodsx. 2020; 7:101065.

[22]

Lekshmi BS, Varanakkottu SN. Janus liquid marbles: fabrication techniques, recent developments, and applications. Droplet. 2023; 2:e44.

[23]

Chen M, Liu Z, Li Y. Magnetic nanofluid-based liquid marble for a self-powered mechanosensation. Droplet. 2024; 3:e122.

[24]

Lathia R, Modak CD, Sen P. Two modes of contact-time reduction in the impact of particle-coated droplets on superhydrophobic surfaces. Droplet. 2023; 2:e89.

[25]

Liu H, Pang X, Duan M, Yang Z, Russell TP, Li X. A simple route for open fluidic devices with particle walls. Adv Mater. 2024:e2413862.

[26]

Wang R, Li X. On the effective surface tension of powder-derived liquid marbles. Powder Technol. 2020; 367: 608-615.

[27]

Li X. Liquid marbles and liquid plasticines with nanoparticle monolayers. Adv Colloid Interface Sci. 2019; 271:101988.

[28]

Lathia R, Nagpal S, Modak CD, et al. Tunable encapsulation of sessile droplets with solid and liquid shells. Nat Commun. 2024; 15: 6445.

[29]

Zhao Z, Yao X, Zhao W, et al. Highly transparent liquid marble in liquid (HT-LMIL) as 3D miniaturized reactor for real-time bio-/chemical assays. Chem Eng J. 2022; 443:136417.

[30]

Li X, Wang Y, Huang J, et al. Monolayer nanoparticle-covered liquid marbles derived from a sol‒gel coating. Appl Phys Lett. 2017; 111:261604.

[31]

Gurav AB, Shi H, Duan M, Pang X, Li X. Highly transparent, hot water and scratch resistant, lubricant-infused slippery surfaces developed from a mechanically-weak superhydrophobic coating. Chem Eng J. 2021; 416:127809.

[32]

Li X, Xue Y, Lv P, et al. Liquid plasticine: controlled deformation and recovery of droplets with interfacial nanoparticle jamming. Soft Matter. 2016; 12: 1655-1662.

[33]

Li X, Shen J. Deforming water droplets with a superhydrophobic silica coating. Chem Commun. 2013; 49: 10016-10018.

[34]

Li X, Shi H, Wang Y, et al. Liquid shaping based on liquid pancakes. Adv Mater Interfaces. 2018; 5:1701139.

[35]

Li X, Wang Y, Wang R, Wang S, Zang D, Geng X. A dip-decoating process for producing transparent bi-superhydrophobic and wrinkled water surfaces. Adv Mater Interfaces. 2018; 5:1800356.

[36]

Boreyko JB. Jumping droplets. Droplet. 2024; 3:e105.

[37]

Li X, Wang R, Shi H, Song B. Effective surface tension of liquid marbles using controllable nanoparticle monolayers. Appl Phys Lett. 2018; 113:101602.

[38]

Shi H, Li X. Monolayer nanoparticle-covered liquid marble production with low surface tension liquids. Adv Mater Interfaces. 2020; 7:2001081.

[39]

Liu H, Peng C, Guo S, Liu X, Li X. Rod-shaped liquid plasticine as cuttable minireactor for photodynamic therapy of tumors. Small. 2024: 20:e2309535.

[40]

Li X, Pang X, Jiang H, et al. Open millifluidics based on powder-encased channels. Proc Natl Acad Sci U S A. 2023; 120:e2302907120.

[41]

Niu J, Liu W, Li JX, et al. Machining water through laser cutting of nanoparticle-encased water pancakes. Nat Commun. 2023; 14: 3853.

[42]

Fujiwara J, Geyer F, Butt H-J, Hirai T, Nakamura Y, Fujii S. Shape-designable polyhedral liquid marbles/plasticines stabilized with polymer plates. Adv Mater Interfaces. 2020; 7:2001573.

[43]

Geyer F, Asaumi Y, Vollmer D, Butt H-J, Nakamura Y, Fujii S. Polyhedral liquid marbles. Adv Funct Mater. 2019; 29:1808826.

[44]

Sun S, Li S, Feng W, Luo J, Russell TP, Shi S. Reconfigurable droplet networks. Nat Commun. 2024; 15: 1058.

[45]

Xia Z, Song Y-F, Shi S. Interfacial preparation of polyoxometalate-based hybrid supramolecular polymers by orthogonal self-assembly. Angew Chem Int Ed Engl. 2024; 63:e202312187.

[46]

Liu X, Kent N, Ceballos A, et al. Reconfigurable ferromagnetic liquid droplets. Science. 2019; 365: 264-267.

[47]

Pang X, Duan M, Liu H, Xi Y, Shi H, Li X. Oscillation-induced mixing advances the functionality of liquid marble microreactors. ACS Appl Mater Inter. 2022; 14: 11999-12009.

[48]

Sun B, Zhao Y, Zhao Q, Li G. A sesseile drop method for facile and robust spheroid cultures. Adv Mater Interfaces. 2021; 8:2100972.

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2025 The Author(s). Droplet published by Jilin University and John Wiley & Sons Australia, Ltd.

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