Pham Hoang Lan, Yang Da Hyun, Chae Woo Ri, Jung Jong Hyeok, Hoang Thi Xoan, Lee Nae Yoon, Kim Jae Young
Department of Life Science, Gachon University, Seongnam 13120, Gyeonggi-Do, Republic of Korea.
Department of BioNano Technology, Gachon University, Seongnam 13120, Gyeonggi-Do, Republic of Korea.
Micromachines (Basel). 2023 Mar 17;14(3):673. doi: 10.3390/mi14030673.
Regulatory macrophages (Mreg) are a special cell type that present a potential therapeutic strategy for various inflammatory diseases. In vitro, Mreg generation mainly takes 7-10 days of treatment with chemicals, including cytokines. In the present study, we established a new approach for Mreg generation using a three-dimensional (3D) micropatterned polydimethylsiloxane (PDMS) surface coated with a natural biopolymer adhesive polydopamine (PDA) and the common cell adhesion peptide motif arginylglycylaspartic acid (RGD). The 3D PDMS surfaces were fabricated by photolithography and soft lithography techniques and were subsequently coated with an RGD+PDA mixture to form a surface that facilitates cell adhesion. Human monocytes (THP-1 cells) were cultured on different types of 2D or 3D micropatterns for four days, and the cell morphology, elongation, and Mreg marker expression were assessed using microscopic and flow cytometric analyses. The cells grown on the PDA+RGD-coated 3D micropatterns (20-µm width/20-µm space) exhibited the most elongated morphology and strongest expression levels of Mreg markers, such as CD163, CD206, CD209, CD274, MER-TK, TREM2, and DHRS9. The present study demonstrated that PDA+RGD-coated 3D PDMS micropatterns successfully induced Mreg-like cells from THP-1 cells within four days without the use of cytokines, suggesting a time- and cost-effective method to generate Mreg-like cells in vitro.
调节性巨噬细胞（Mreg）是一种特殊的细胞类型，为各种炎症性疾病提供了一种潜在的治疗策略。在体外，生成Mreg主要需要用包括细胞因子在内的化学物质处理7 - 10天。在本研究中，我们建立了一种生成Mreg的新方法，使用涂有天然生物聚合物粘合剂聚多巴胺（PDA）和常见细胞粘附肽基序精氨酰甘氨酰天冬氨酸（RGD）的三维（3D）微图案化聚二甲基硅氧烷（PDMS）表面。3D PDMS表面通过光刻和软光刻技术制造，随后用RGD + PDA混合物包被以形成促进细胞粘附的表面。将人单核细胞（THP - 1细胞）在不同类型的二维或三维微图案上培养四天，使用显微镜和流式细胞术分析评估细胞形态、伸长情况和Mreg标志物表达。在涂有PDA + RGD的3D微图案（宽度20 µm/间距20 µm）上生长的细胞表现出最细长的形态以及最强的Mreg标志物表达水平，如CD163、CD206、CD209、CD274、MER - TK、TREM2和DHRS9。本研究表明，涂有PDA + RGD的3D PDMS微图案在不使用细胞因子的情况下，四天内成功从THP - 1细胞诱导出Mreg样细胞，提示了一种在体外生成Mreg样细胞的省时且经济高效的方法。
