Zhao Yajun, Xiao E, Lv Wanqi, Dong Xian, He Linhai, Wang Yixiang, Zhang Yi
Department of Oral and Maxillofacial Surgery, Peking University School and Hospital of Stomatology, Beijing 100081, China.
Peking University Hospital of Stomatology First Clinical Division, Beijing 100081, China.
Stem Cells Int. 2020 Mar 7;2020:1031985. doi: 10.1155/2020/1031985. eCollection 2020.
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 and . After transplantation into mouse, spheroid ADSCs improved the survival rate and significantly decreased serum levels of proinflammatory factors IL-1 and TNF- 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.
间充质干细胞（MSCs）在组织工程和再生医学中具有广阔的应用潜力。先前的研究表明，与单层培养相比，MSCs形成球体表现出更好的干性维持和治疗潜力。迄今为止，已经开发了各种球体培养系统，但大多数都需要低粘附条件或特殊设备。在本研究中，我们证明在TeSR-E8培养基中接种解离的MSCs可以在传统的组织培养聚苯乙烯培养皿中诱导自组装球体形成。与单层培养相比，脂肪来源干细胞（ADSC）球体增强了ADSCs的增殖和成骨能力。当重新接种到含正常血清的培养基中时，球体来源的ADSCs中干性生物标志物的表达水平甚至高于单层培养。重要的是，球体ADSCs在体内和体外都能有效促进巨噬细胞的M2极化。移植到小鼠体内后，球体ADSCs提高了存活率，并在脂多糖攻击后显著降低了促炎因子IL-1和TNF的血清水平。总之，我们通过TeSR-E8培养基开发了一种三维球体培养系统，无需低粘附条件和特殊设备的参与，为MSCs球体形成提供了一种实用且便捷的方法，在干细胞临床治疗中具有巨大潜力。
