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.
Highlights
Mesenchymal stem cells (MSCs) possess tremendous potential for tissue engineering and regenerative medicine due to their strong self-renewal, multipotent differentiation potential, and the immunomodulatory capacity [1]
Our study demonstrated that MSCs cultured in regular tissue culture dishes could spontaneously form 3D spheroids in a chemically defined serum-free culture condition without the involvement of low adhesion conditions and special equipment, which provided a convenient method for spheroid formation of MSCs with the maintenance of stemness and enhancement of their therapeutic potential
To verify whether MSCs from other sources have the same characteristics as adipose-derived stem cell (ADSC), two types of MSCs, Bone marrow mesenchymal stem cells (BMSCs) and Dental pulp stem cells (DPSCs), were cultured in TeSR-E8 medium
Summary
MSCs possess tremendous potential for tissue engineering and regenerative medicine due to their strong self-renewal, multipotent differentiation potential, and the immunomodulatory capacity [1]. The most important thing is how to maintain the stemness and the therapeutic potential of MSCs. Traditionally, the standard method for in vitro expansion of MSCs is two-dimensional (2D) adherent culture. MSCs are usually subjected to impaired stemness and multipotency including a loss of proliferation rate and differentiation capacity during long-time, monolayer culture, which greatly reduced the therapeutic efficacy [4]. Many studies have demonstrated that MSCs cultured in 3D spheroids showed enhanced viability, stemness, and differentiation potential compared to monolayer cells [6, 12, 13]. MSC spheroids have showed improved therapeutic efficacy in preclinical animal studies, such as bone and skin defect [14, 15], Stem Cells International ischemic disease [16], and cardiovascular disorders [16, 17]. The application of 3D spheroid cell culture techniques is receiving an increased attention
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