Recent Advances in Three-Dimensional Stem Cell Culture Systems and Applications

Abstract

Cell culture is one of the most core and fundamental techniques employed in the fields of biology and medicine. At present, although the two-dimensional cell culture method is commonly used in vitro, it is quite different from the cell growth microenvironment in vivo. In recent years, the limitations of two-dimensional culture and the advantages of three-dimensional culture have increasingly attracted more and more attentions. Compared to two-dimensional culture, three-dimensional culture system is better to realistically simulate the local microenvironment of cells, promote the exchange of information among cells and the extracellular matrix (ECM), and retain the original biological characteristics of stem cells. In this review, we first present three-dimensional cell culture methods from two aspects: a scaffold-free culture system and a scaffold-based culture system. The culture method and cell characterizations will be summarized. Then the application of three-dimensional cell culture system is further explored, such as in the fields of drug screening, organoids and assembloids. Finally, the directions for future research of three-dimensional cell culture are stated briefly.

Application of Hexanoyl Glycol Chitosan as a Non-cell Adhesive Polymer in Three-Dimensional Cell Culture

“… 26 Various methods such as rotating bioreactor culture and magnetic suspension culture have been introduced to prevent spheroid fusion and to independently culture spheroids for a long time. 27 However, an ideal solution for uncontrollable fusion of spheroids has not yet been reported. Although spheroids in the control group were fused, the treatment of the soluble HGC improved the 3D scaffold-free culture, maintaining individual spheroids during 14 days of culture by inhibiting the adhesive property of cells as we hypothesized.…”

Strategies to Modulate the Redifferentiation of Chondrocytes

“…Gelatin-methacryloyl hydrogels, tyrosinase-crosslinked alginate sulfate tyramine hydrogels, and composite microfibers also exhibit potential for the engineering of cartilage-like tissues for 3D cultured chondrocytes (Klotz et al, 2016;Angelozzi et al, 2017;Öztürk et al, 2020). Self-assembled dendritic DNA hydrogel possesses inherent biocompatibility, biodegradability, and unique programmability, and has shown great potential in three- dimensional cell culture (Wu J. et al, 2021), but its effect on the redifferentiation of chondrocytes needs further investigation.…”

“…At present, 3D culture is widely used in cell culture, because researchers increasingly find that the extracellular environment of 2D culture is far from the real extracellular microenvironment. 3D culture can be classified into 3D culture methods with and without scaffolds ( Wu X. et al, 2021 ), and Three-dimensional culture with scaffolds is widely used to redifferentiate chondrocytes ( Caron et al, 2012 ). This type of culture is a common approach that is used to induce and maintain chondrocyte redifferentiation, and generate ECM.…”

Formation of Three-Dimensional Spheres Enhances the Neurogenic Potential of Stem Cells from Apical Papilla

“…However, longterm in-vitro culture and serial passages by using the conventional two-dimensional (2D) dish, are quite likely to evoke continuous changes in MSCs. Stem cells may gradually lose self-renewal ability, differentiate into bone cells or adipocytes spontaneously, and undergo senescence [11]. To overcome these deleterious effects and their possible consequent functional alterations, 3D cell culture has been employed in various studies, which can better simulate the growth state of cells in-vivo, promote cell self-renewal, inhibit their differentiation, and maintain the original characteristics of cells [11,12].…”

“…Stem cells may gradually lose self-renewal ability, differentiate into bone cells or adipocytes spontaneously, and undergo senescence [11]. To overcome these deleterious effects and their possible consequent functional alterations, 3D cell culture has been employed in various studies, which can better simulate the growth state of cells in-vivo, promote cell self-renewal, inhibit their differentiation, and maintain the original characteristics of cells [11,12]. For example, short-term spheroid formation of adipose-derived stem cells (ASCs) can enhance their stemness, angiogenic capacity, and chemotaxis and thereby increase their therapeutic potential for tissue regeneration [13].…”