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Cell Patterning Using Magnetic-Archimedes Strategy
In This Article
Summary
This protocol describes an ink-free, label-free, substrate-independent, high-throughput cell patterning method based on the Magnetic Archimedes effect.
Abstract
Cell patterning, allowing precise control of cell positioning, presents a unique advantage in the study of cell behavior. In this protocol, a cell patterning strategy based on the Magnetic-Archimedes (Mag-Arch) effect is introduced. This approach enables precise control of cell distribution without the use of ink materials or labeling particles. By introducing a paramagnetic reagent to enhance the magnetic susceptibility of the cell culture medium, cells are repelled by magnets and arrange themselves into a pattern complementary to the magnet sets positioned beneath the microfluidic substrate.
In this article, detailed procedures for cell patterning using the Mag-Arch-based strategy are provided. Methods for patterning single-cell types as well as multiple cell types for co-culture experiments are offered. Additionally, comprehensive instructions for fabricating microfluidic devices containing channels for cell patterning are provided. Achieving this feature using parallel methods is challenging but can be done in a simplified and cost-effective manner. Employing Mag-Arch-based cell patterning equips researchers with a powerful tool for in vitro research.
Introduction
Cell patterning is evolving into an intuitive and powerful technology for in vitro studies1. By manipulating cell positions in culture plates, it provides solutions for a variety of experiments, including cell migration2, biomimetic multicellular co-culture3, organoid assembly4, biomaterial studies5, and more. In most situations, an ink-free, label-free method is preferred for cell patterning because it offers ease of operation and high cell viability for subsequent investigations.
The Mag-Arch effect is a physica....
Protocol
1. Assembling the magnet sets
- Assemble the magnet sets for strip patterns.
- Choose flat rectangular magnets, as depicted in Figure 1A. The dimensions of the rectangular magnets used for this demonstration are 1.5 mm × 10 mm × 35 mm (thickness × height × length) (see Table of Materials). The thickness of the magnets determines the gaps between the cell stripes.
- Cut 2 mm-thick silicone plates (see T.......
Representative Results
Rectangular (1.5 mm × 10 mm × 35 mm) and cylindrical (Φ1.5 m × 10 mm) magnets were selected to create cell patterns as a demonstration. Users have the flexibility to modify the size and shape of magnets or assemble them differently to create diverse cell patterns. In Figure 1A,B, the magnets were assembled, with the magnetic poles depicted in blue (south) and red (north) for clarity. In this configuration, magnets attract each other laterally and align th.......
Discussion
The Mag-Arch-based cell patterning provides a user-friendly solution for most biomedical laboratories. This method advances parallel to characters of ink-free, label-free, substrate-independent, and the ability for high-throughput patterning8,13. For mono-type cell patterning, it patterns cells in a one-step way. The procedure finishes simply by refreshing culture mediums.
Previous studies have used magnetic particles to label cells an.......
Acknowledgements
This study is financially supported by the National Key R&D Program of China (2021YFA1101100), the National Natural Science Foundation of China (32000971), the Fundamental Research Funds for the Central Universities (No. 2021FZZX001-42), and the Starry Night Science Fund of Zhejiang University Shanghai Institute for Advanced Study (Grant No. SN-ZJU-SIAS-004).
....Materials
| Name | Company | Catalog Number | Comments |
| A2780 ovarian cancer cells | Procell | CL-0013 | |
| Cell culture medium (DMEM, high glucose) | Gibco | 11995040 | |
| Cover slides | Citotest Scientific | 80340-3610 | For fabricating microfluidics. Dimension: 24 mm × 50 mm |
| DiD | MedChemExpress (MCE) | HY-D1028 | For labeling cells with red fluorescence (Ex: 640 nm) |
| DiI | MedChemExpress (MCE) | HY-D0083 | For labeling cells with orange fluorescence (Ex: 550 nm) |
| Fetal Bovine Serum (FBS) | Biochannel | BC-SE-FBS07 | |
| Gadopentetate dimeglumine (Gd-DTPA) | Beijing Beilu Pharmaceutical | H10860002 | |
| Gelatin | Sigma Aldrich | V900863 | |
| Glass cell slides | Citotest Scientific | 80346-2510 | Diameter: 25 mm; thickness: 0.19-0.22 mm |
| Glass plates | PURESHI hardware store | For fabricating microfluidics. Dimension: 40 mm × 75 mm | |
| Human Umbilical Vein Endothelial Cells (HUVECs) | Servicebio | STCC12103G-1 | |
| Neodymium-iron-boron magnets (N52) | Lalaci | ||
| Non-toxic glass plate coating (Gel Slick Solution) | Lonza | 1049286 | For convenience of demolding when fabricating microfluidics |
| Phosphate Buffered Saline (PBS) | Servicebio | G4200 | |
| Plasma cleaner | SANHOPTT | PT-2S | |
| Polydimethylsiloxane (PDMS) kit | DOWSIL | SYLGARD 184 Silicone Elastomer Kit | For fabricating microfluidics |
| Polytetrafluoroethylene (PFTE) mold | PURESHI hardware store | Customized online, for fabricating microfluidics | |
| Silicon plate | PURESHI hardware store | ||
| Smooth Muscle Cells (SMC) | Procell | CL-0517 | |
| Ultrasonic cleaner | Sapeen | CSA-02 |
References
- Christian, J., et al. Control of cell adhesion using hydrogel patterning techniques for applications in traction force microscopy. J Vis Exp. 179, e63121 (2022).
- Abbas, Y., Turco, M. Y., Burton, G. J., Moffett, A.
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