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A rapid and quantitative method for the analysis of cell migration using retroreflective Janus particular probe and micropatterned surface

  • 양은경 (아주대학교 일반대학원)

초록/요약

Cell migration is a fundamental process that plays a crucial role in various physiological and pathological events. By understanding the factors influencing cell migration, we can advance scientific knowledge and develop potential therapeutic interventions. Consequently, there is a growing demand for reliable and versatile cell migration assays that can be used for in vitro analysis. The scratch wound healing and Transwell assays are conventional methods for studying cell migration. However, they have limitations such as manual wound creation and the inability to track individual cell movements in real-time. Herein, we developed a novel assay that combines the advantages of these methods. This assay utilizes a micropatterned plate, eliminating the need for manual scratches and incorporating a retroreflection optical analysis system for precise cell tracking. By introducing a retroreflective system with micro-scale mirror particles, we achieved real-time cell migration tracking with minimal interference from the surrounding environment. The assay performance was evaluated using L929 fibroblasts treated with cyclic Arg-Gly-Asp as a migratory control molecule, and the degree of cell migration was quantified by analyzing the positional change of retroreflective signals. The results demonstrated the feasibility of the developed assay for analyzing cell migration. This novel method provides a tool for quantitative and controlled cell migration analysis, facilitating further research and potential therapeutic interventions.

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목차

1. Introduction 1
1.1 Significance of studying the cell migration phenomenon 1
1.2 Standard method for analyzing in vitro cell migration 2
1.3 Novel cell migration assay using retroreflective Janus particles and micropatterned plate 3
1.4 Sensing strategy for quantification of cell mobility 5
1.5 Aim of Thesis 8
2. Materials and methods 9
2.1 Reagents and apparatus 9
2.2 Treatment of cell migration inhibitory materials on the patterned surface 10
2.2.1 Confirmation of migration inhibition by different structural of RGD 10
2.2.2 Confirmation of migration inhibition by cyclic RGD 11
2.3 Preparation of optical probe 11
2.4 Cell viability test 12
2.5 Analysis of cell mobility using the developed system 13
2.5.1 Quantification of cell mobility treated with cyclic RGD 13
2.5.2 Quantification of cell mobility treated with various substances 13
3. Results and discussion 14
3.1 Working principle of developed quantitative cell migration assay 14
3.2 Verification of the designed cell culture substrate 15
3.2.1 Fabrication of micropattern plate 15
3.2.2 Observation of cell migration on the pattern 18
3.3 Verification of applicability of reagents and optical probe for quantitative analysis of cell migration 21
3.3.1 Observation of cell migratory control effect by different structural of RGD 21
3.3.2 Observation of cell migratory control effect by cyclic RGD 24
3.3.3 Confirmation of functionalization of the RJPs 26
3.3.4 Confirmation of cytotoxicity of RGD and RJPs 29
3.4 Quantitative analysis of cell mobility 31
3.4.1 Analysis methodology development of retroreflection-based cell migration assay 31
3.4.2 Quantification of cell migration inhibitory effects using RJPs 37
3.4.3 Application of the developed sensing system to various substances 41
4. Conclusions 43
5. References 45

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