Overexpression of α₅β₁ Integrin in HEK293T Cell Enhances Cell–Cell and Cell–ECM Interactions Enabling Multi-Cell Layer Fabrication
- 주제(키워드) multi-cell layer , intercellular cohesion , α5β1 integrin
- 발행기관 서강대학교 일반대학원
- 지도교수 신관우
- 발행년도 2026
- 학위수여년월 2026. 2
- 학위명 석사
- 학과 및 전공 일반대학원 화학과
- 실제URI http://www.dcollection.net/handler/sogang/000000082598
- UCI I804:11029-000000082598
- 본문언어 영어
- 저작권 논문은 저작권에 의해 보호받습니다.
초록(요약문)
Cell–cell and cell–extracellular matrix (ECM) interactions are essential for maintaining the structural and functional integrity of tissues. The integrin receptor α₅β, a principal fibronectin-binding molecule, plays a pivotal role in mediating these adhesive interactions. In this study, we investigated how overexpression of α₅β integrin enhances intercellular and cell–ECM adhesion using human embryonic kidney (HEK293T) cells as a model system. HEK293T cells were genetically modified to overexpress α₅β₁ integrin via viral transduction, followed by selection using fluorescence-activated cell sorting (FACS). Successful overexpression was confirmed by flow cytometry analysis, western blot and immunocytostaining. Functional assays demonstrated that α₅β₁integrin overexpression significantly enhanced cell aggregation and cell–ECM adhesion properties compared to control cells. Furthermore, as an application of enhanced adhesion, α₅β₁integrin-overexpressed cells exhibited improved structural organization in multi-cell layer assembly, suggesting their potential utility in scaffold-free tissue engineering approaches. These findings highlight the critical role of α₅β₁integrin in regulating adhesive properties and its promise as a molecular target for advancing cell-based tissue construction.
more목차
1. Introduction 1
2. Experimental details 4
2.1. Cell culture 4
2.2. Integrin α5 gene Transduction 4
2.3. FACS and flow cytometry analysis 5
2.4. Western blot analysis 5
2.5. Immunofluorescent staining 6
2.6. In vitro experiments 7
2.7. Fabrication of multi-cell layers 7
2.8. Statistical analysis 7
3. Results and discussion 9
3.1. Establishment of integrin α5β1-overexpressed HEK293T cells by
lentiviral transduction 9
3.2. Verification of integrin α5β1 overexpression at the cell-surface and
total protein levels 11
3.3. Enhancement of cell–ECM adhesion and cell–cell aggregation by integrin α5β1 overexpression 13
3.4. Fabrication of 3D multi-cell layers using integrin α5β1-overexpressed cells 19
4. Conclusion 23
References 26
