THREE-DIMENSIONAL CELL CULTURES IN BIOTECHNOLOGY: A CONTEMPORARY PERSPECTIVE Trokhimenko O. P., Mysko Yu. P., Soloviov S. O., Rossokha Z .I., Smetiukh M. P., Trokhymchuk V. V., Pryputa N. V., Rymar M. V., Leseiko T. O.

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SSN 2410-7751 (Print)
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cover biotech acta general

Biotechnologia Acta Т. 19, No. 4, 2026
P. 5-18 , Bibliography 56, Engl.
UDC 577.3:602.9:57.085.23
doi: https://doi.org/10.15407biotech19.04.005

Full text: (PDF, in English)

THREE-DIMENSIONAL CELL CULTURES IN BIOTECHNOLOGY: A CONTEMPORARY PERSPECTIVE

¹ Shupyk National Healthcare University of Ukraine, Kyiv, Ukraine
² National Technical University of Ukraine “Igor Sikorsky Kyiv Polytechnic Institute”, Kyiv, Ukraine

Aim. To systematize and critically analyze contemporary three-dimensional (3D) cell culture platforms — spheroids, organoids, bioprinted constructs, and organs-on-chips — and to evaluate their potential as physiologically relevant alternatives to two-dimensional (2D) monolayer systems in biomedical research and preclinical drug evaluation.

Methods. A narrative review of current scientific literature was conducted, encompassing the principal categories of 3D cell culture technologies, their underlying biological and engineering principles, cultivation methodologies, extracellular matrix (ECM) composition, and available standardization platforms. Comparative analysis of 2D and 3D culture properties was performed, alongside evaluation of key analytical approaches — including Ki-67 immunolabeling, BrdU/EdU incorporation, metabolic assays, and confocal microscopy — adapted for three-dimensional structures.

Results. Three-dimensional cell cultures substantially surpass 2D monolayers in recapitulating cell–cell and cell–ECM interactions, diffusion gradients, and tissue-specific spatial organization. Each principal platform demonstrates distinct applicability: MCTS reproduces solid tumor zonal architecture for oncological and pharmacological studies; organoids enable tissue-specific self-organization for disease modeling and personalized medicine; bioprinting allows precise spatial deposition of cells and scaffolds; and organs-on-chips integrate microfluidic perfusion to model dynamic physiological barrier functions.

Conclusions. Three-dimensional cell culture systems represent a scientifically substantiated and ethically consistent alternative to animal models, advancing the implementation of the 3Rs principles in experimental biomedicine. Nevertheless, their broad translational application remains constrained by insufficient standardization, limited reproducibility, high operational costs, and the absence of functional vascularization. Overcoming these barriers requires harmonizing validated protocols and integrating them with high-throughput analytical platforms.

Keywords: three-dimensional cell cultures, spheroids, organoids, organs-on-chips, bioprinting, extracellular matrix, bioinks, oncology, pharmacology, regenerative medicine.

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