A Chemically Defined Serum-Free Culture System for Spontaneous Human Mesenchymal Stem Cell Spheroid Formation

Abstract

Mesenchymal stem cells (MSCs) possess promising potential in tissue engineering and regenerative medicine. Previous studies demonstrated that spheroid formation of MSCs exhibited improved stemness maintenance and therapeutic potential compared with monolayer culture. To date, various spheroid culture systems have been developed but most of them required low adhesion conditions or special equipment. In this study, we demonstrated that inoculation of dissociated MSCs in TeSR-E8 medium could induce self-assemble spheroid formation in conventional tissue culture polystyrene dishes. Compared with monolayer culture, adipose-derived stem cell (ADSC) spheroids enhanced the proliferation and osteogenic capability of ADSCs compared with monolayer culture. When reseeded in normal serum-containing medium, the expression level of stemness biomarkers was even higher in spheroid-derived ADSCs than monolayer culture. Importantly, spheroid ADSCs could effectively promote the M2 polarization of macrophages both in vitro and in vivo. After transplantation into mouse, spheroid ADSCs improved the survival rate and significantly decreased serum levels of proinflammatory factors IL-1β and TNF-α following LPS challenge. In summary, we developed a 3D spheroid culture system through TeSR-E8 medium without the involvement of low adhesion conditions and special equipment, which provided a practical and convenient method for spheroid formation of MSCs with great potential for stem cell clinical therapy.

In Vitro and In Vivo Characterization of PLLA-316L Stainless Steel Electromechanical Devices for Bone Tissue Engineering—A Preliminary Study

“…Small differences were observed regarding cell appearance, probably due to the topographic differences between both devices. Differentiated cells presented some extent of spontaneous cell aggregation, a behavior previously described for MSCs cultured in low serum supplemented medium, and thus considered normal [ 31 ]. This phenomenon can also be linked to an increase potential of these cells to differentiate into the osteogenic lineage, as aggregates have been widely reported to present superior differentiation potential compared to the 2D cell culture [ 32 , 33 ].…”

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

“…Zhao et al have verified that human mesenchymal stem cells with a chemically defined serum-free TeSR-E8 medium can spontaneously assemble into 3D spheres. And the research found compared to 2D, the stemness of 3D cells is enhanced, which increases the treatment efficiency of endotoxemia mice and reduces mortality [ 6 ].…”

“…For example, stem cells are prone to lose self-renewal ability, become senescence, or differentiate into bone cells or adipocytes spontaneously [ 5 ]. Long-term cultures will gradually lead to tissue specificity lost [ 6 ], observed differences between the results of in vitro cell culture and in vivo animal experiments. Therefore, understanding how to better simulate the physiological environment under in vitro conditions is very important for medical research [ 7 ].…”

Human iPSC-Vascular Smooth Muscle Cell Spheroids Demonstrate Size-dependent Alterations in Cellular Viability and Secretory Function

“…13,16,17 In comparison to the traditional culture technique of cells in a monolayer, cells cultured as spheroids have demonstrated enhanced cell-survival, paracrine factor secretion, and tissue forming potential. 3,4,6,8,9,[18][19][20][21] They also have higher levels of tissue-specific gene expression and increased exosome production. 10,22 Overall, spheroids enhance biological processes relating to immunity, inflammation, angiogenesis, and wound healing.…”