Citation: Huang, X.; Huang, Z.; Gao, W.; Gao, W.; He, R.; Li, Y.; Crawford, R.; Zhou, Y.; Xiao, L.; Xiao, Y. Current Advances in 3D Dynamic Cell Culture Systems. Gels 2022, 8, 829. https://doi.org/10.3390/gels8120829 Academic Editor: Tal Dvir Received: 1 November 2022 Accepted: 13 December 2022 Published: 16 December 2022 Publisher’s Note: MDPI stays neutral with regard to jurisdictional claims in published maps and institutional affil- iations. Copyright: © 2022 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https:// creativecommons.org/licenses/by/ 4.0/). gels Review Current Advances in 3D Dynamic Cell Culture Systems Xin Huang 1,† , Zhengxiang Huang 1,2,† , Weidong Gao 1,† , Wendong Gao 1 , Ruiying He 3 , Yulin Li 4 , Ross Crawford 1,5 , Yinghong Zhou 5,6 , Lan Xiao 1,5, * and Yin Xiao 1,5,7, * 1 School of Mechanical, Medical and Process Engineering, Center of Biomedical Technology, Queensland University of Technology, Brisbane, QLD 4059, Australia 2 School of Biomedical Sciences, The University of Queensland, St Lucia, QLD 4072, Australia 3 College of Chemistry and Chemical Engineering, Hubei University, Wuhan 430062, China 4 The Key Laboratory for Ultrafine Materials of Ministry of Education, State Key Laboratory of Bioreactor Engineering, Engineering Research Center for Biomedical Materials of Ministry of Education, School of Materials Science and Engineering, East China University of Science and Technology, Shanghai 200237, China 5 Australia-China Centre for Tissue Engineering and Regenerative Medicine, Queensland University of Technology, Brisbane, QLD 4059, Australia 6 School of Dentistry, The University of Queensland, Herston, QLD 4072, Australia 7 School of Medicine and Dentistry, Griffith University, Gold Coast, QLD 4222, Australia * Correspondence: l5.xiao@qut.edu.au (L.X.); yin.xiao@qut.edu.au or yin.xiao@griffith.edu.au (Y.X.) † These authors contributed equally to this work. Abstract: The traditional two-dimensional (2D) cell culture methods have a long history of mimicking in vivo cell growth. However, these methods cannot fully represent physiological conditions, which lack two major indexes of the in vivo environment; one is a three-dimensional 3D cell environment, and the other is mechanical stimulation; therefore, they are incapable of replicating the essential cellular communications between cell to cell, cell to the extracellular matrix, and cellular responses to dynamic mechanical stimulation in a physiological condition of body movement and blood flow. To solve these problems and challenges, 3D cell carriers have been gradually developed to provide a 3D matrix-like structure for cell attachment, proliferation, differentiation, and communication in static and dynamic culture conditions. 3D cell carriers in dynamic culture systems could primarily provide different mechanical stimulations which further mimic the real in vivo microenvironment. In this review, the current advances in 3D dynamic cell culture approaches have been introduced, with their advantages and disadvantages being discussed in comparison to traditional 2D cell culture in static conditions. Keywords: 3D cell culture; mechanical stimulation on cell behavior; bioreactor; microcarrier; organ- on-a-chip 1. Introduction Since Harrison Ross first carried out in vitro cell culture using a sterile coverslip in 1906 [1], the era for cell culture began. Nowadays, the cell culture technique is one of the most common techniques in many fields of biomedical sciences, from basic research to large-scale industrial production of biological products. It offers an efficient approach to achieving different purposes without using animals. To culture most of the cell types outside of a living body, artificial devices are usually required to allow the cells to adhere and grow. Glass devices such as coverslips were most commonly used in the first few decades of cell culture history [2]. Later, plasma- treated polystyrene was invented by the Falcon Plastics Company and showed excellent properties for cell adhesion and growth [3]. More recently, plasma-treated polystyrene has dominated the research consumer market with different configurations designed for various research purposes, such as flasks, dishes, and plates. These cell culture devices allow adherent cells to grow in a monolayer on a two-dimensional (2D) planar surface under static Gels 2022, 8, 829. https://doi.org/10.3390/gels8120829 https://www.mdpi.com/journal/gels