Yao Chao-Ling, Tseng Tsung-Yu
Department of Chemical Engineering, National Cheng Kung University, No. 1, University Road, Tainan, 70101, Taiwan.
J Biol Eng. 2023 Jan 26;17(1):6. doi: 10.1186/s13036-023-00325-z.
Dendritic cells (DCs) are differentiated from monocytes, and have a strong ability to perform phagocytosis, present antigens and activate T cell immune response. Therefore, DCs are one of the key factors in fighting cancer in immunotherapy, and it is an important issue to develop a serum-free system for DC differentiation and expansion in vitro for clinical application.
In this study, IL-6 and M-CSF were determined and a concentration combination of cytokines was optimized to develop an optimal DC serum-free differentiation medium (SF-DC Optimal) that can effectively differentiate CD14 monocytes into CD40CD209 DCs. After differentiation, the morphology, growth kinetics, surface antigen expression, phagocytosis ability, cytokine secretion, mixed lymphocyte reaction and stimulation for maturation of the differentiated DCs were checked and confirmed. Importantly, this research is the first report finding that the addition an extra low concentration of IL-6 and M-CSF exhibited a synergistic effect with GM-CSF and IL-4 to generate higher numbers and more fully functional DCs than the addition of GM-CSF and IL-4 only under serum-free condition.
A large number of functional DCs can be generated by using SF-DC Optimal medium and provide an alternative source of DCs for related basic research and clinical applications.
树突状细胞（DCs）由单核细胞分化而来，具有很强的吞噬、呈递抗原及激活T细胞免疫反应的能力。因此，DCs是免疫治疗中对抗癌症的关键因素之一，开发用于DCs体外分化和扩增的无血清体系以用于临床应用是一个重要问题。
在本研究中，测定了白细胞介素-6（IL-6）和巨噬细胞集落刺激因子（M-CSF），并优化了细胞因子的浓度组合，以开发一种最佳的DCs无血清分化培养基（SF-DC Optimal），该培养基能有效地将CD14单核细胞分化为CD40CD209 DCs。分化后，对分化后的DCs的形态、生长动力学、表面抗原表达、吞噬能力、细胞因子分泌、混合淋巴细胞反应及成熟刺激进行了检测和确认。重要的是，本研究首次报道发现在无血清条件下，添加极低浓度的IL-6和M-CSF与粒细胞-巨噬细胞集落刺激因子（GM-CSF）和白细胞介素-4（IL-4）具有协同作用，能产生比仅添加GM-CSF和IL-4数量更多、功能更完备的DCs。
使用SF-DC Optimal培养基可产生大量功能性DCs，为相关基础研究和临床应用提供了DCs的替代来源。
