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Updated: Aug 5, 2025
Production and Characterization of Human Macrophages from Pluripotent Stem Cells
Published on: April 16, 2020
PDMS Micropatterns Coated with PDA and RGD Induce a Regulatory Macrophage-like Phenotype
Hoang Lan Pham1, Da Hyun Yang2, Woo Ri Chae2
1Department of Life Science, Gachon University, Seongnam 13120, Gyeonggi-Do, Republic of Korea.
Abstract:
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.
Insights
This study introduces a novel method for generating regulatory macrophages (Mreg) using 3D micropatterned surfaces. This approach accelerates Mreg production in vitro within four days, offering a faster and more cost-effective alternative.
Area of Science:
- Biomaterials Science
- Cell Biology
- Immunology
Background:
- Regulatory macrophages (Mreg) are crucial for managing inflammatory diseases.
- Current in vitro Mreg generation methods are lengthy, requiring 7-10 days and cytokine treatment.
Purpose of the Study:
- To develop a rapid and efficient method for generating Mreg-like cells in vitro.
- To investigate the use of 3D micropatterned polydimethylsiloxane (PDMS) surfaces coated with polydopamine (PDA) and arginylglycylaspartic acid (RGD) for Mreg generation.
Main Methods:
- Fabrication of 3D PDMS micropatterns using photolithography and soft lithography.
- Coating of PDMS surfaces with a PDA and RGD mixture to enhance cell adhesion.
- Culture of human monocytes (THP-1 cells) on various 2D and 3D micropatterned surfaces for four days.
- Assessment of cell morphology, elongation, and Mreg marker expression via microscopy and flow cytometry.
Main Results:
- THP-1 cells cultured on PDA+RGD-coated 3D micropatterns (20-µm width/20-µm space) showed enhanced elongated morphology.
- These cells exhibited significantly higher expression of key Mreg markers (CD163, CD206, CD209, CD274, MER-TK, TREM2, DHRS9).
- Mreg-like cells were successfully generated within four days without cytokine induction.
Conclusions:
- PDA+RGD-coated 3D PDMS micropatterns provide an effective platform for rapid in vitro Mreg generation.
- This novel approach offers a time- and cost-efficient alternative to conventional methods for producing Mreg-like cells.
